The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large scale, with an improved yield and/or with an improved efficiency. The species of cation present in the nucleotide salt as the counter-ion is critical to the yield, efficiency and fidelity of high yielding enzymatic DNA synthesis reaction. The processes herein use alternative cations as counter-ions for the ionic nucleotides, permitting the use of higher concentrations of nucleotides in DNA synthesis, and further allowing for more favourable reaction conditions to be used.
The present invention relates to an improvement to cell-free, in vitro processes that involve the generation of single-stranded nucleic acid concatemers, where these concatemers favour folding into secondary structures due to the presence of stretches of self-complementary sequence. Single stranded concatemers may be produced via rolling circle amplification of a template nucleic acid under certain conditions. The self-annealing and folding of these single-stranded concatemers can frustrate the use of these concatemers for further isolation or product production, particularly where it is desired to make double stranded DNA. The present invention addresses this need by providing a simple and elegant way to ensure that the single stranded nucleic acid concatemer can effectively be duplexed, even if it forms complex self-annealed structures.
B82B 1/00 - Nanostructures formées par manipulation d’atomes ou de molécules, ou d’ensembles limités d’atomes ou de molécules un à un comme des unités individuelles
The present invention relates to novel nucleotide complexes which enhance the enzymatic production of DNA. The nucleotide complexes includes a divalent cation at a ratio of 0.2-1.5 divalent cations per nucleotide, whilst monovalent ions (other than hydrogen or hydronium ions from any solvent) are minimal. The nucleotide complexes have desirable properties for the synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale. Further, the invention includes improved processes for preparing said nucleotide complexes. Further complexes are disclosed which possess unique properties in accelerating DNA synthesis at higher concentrations, these complexes also containing zwitterion molecules.
The present invention relates to a novel closed linear DNA vector, which is suitable for use in the production of lentiviral particles. Notably, the present invention relates to a new configuration of the vector including the transgene (often termed the “payload” vector), which enables a greater yield of infectious lentiviral particles, notably a greater yield of lentiviral particles carrying a trans-gene, to be prepared when compared to closed linear DNA vectors lacking this configuration. Further, the inventors have developed improvements in lentiviral production with closed linear DNA, through optimisation of vector input quantities and construct ratios. The invention furthermore relates to a method of generating infectious lentiviral particles using the construct, optionally in conjunction with improved production vectors and/or optimised methodology.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals, enzymes, nucleic acid constructs and viral vectors for scientific and research use; nucleic acid constructs and viral vectors for use in the manufacture of nucleic acids; nucleic acid constructs and viral vectors for manufacturing viruses; nucleic acid constructs and viral vectors for use in scientific research, agribiotechnology, and nanotechnology; all of the aforementioned goods not for medical or veterinary use Nucleic acid constructs for medical and veterinary purposes; nucleic acid constructs and viral vectors for use as prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for the production of prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for medical use; nucleic acid constructs and viral vectors for the production of therapeutic or prophylactic viruses; nucleic acid constructs and viral vectors for gene therapy; nucleic acid constructs and viral vectors for use in cell therapy; nucleic acid constructs and viral vectors for use in genome editing; nucleic acid constructs and viral vectors for use in therapy Scientific and technological services, namely, research and design in the fields of chemicals, enzymes, nucleic acid constructs, and viral vectors; chemical and biological analysis and research services; research into nucleic acids and nucleic acid constructs; research and development into the creation of nucleic acid constructs; research, analysis, design, and development relating to therapies, prophylactics, and counteragents based on nucleic acid constructs; research and development in the fields of gene therapy, diagnostic preparation, agribiotechnology, and nanotechnology; consultancy relating to the aforesaid services
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
40 - Traitement de matériaux; recyclage, purification de l'air et traitement de l'eau
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals, enzymes, nucleic acid constructs and viral vectors for scientific and research use; nucleic acid constructs and viral vectors for use in the manufacture of nucleic acids; nucleic acid constructs and viral vectors for manufacturing viruses; nucleic acid constructs and viral vectors for use in scientific research, agribiotechnology, and nanotechnology; two-dimensional and three-dimensional nucleic acid constructs for creating nanostructures and scaffolds; all of the aforementioned goods not for medical or veterinary use Nucleic acid constructs for medical and veterinary purposes; nucleic acid constructs and viral vectors for use as prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for the production of prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for medical use; nucleic acid constructs and viral vectors for the production of therapeutic or prophylactic viruses; nucleic acid constructs and viral vectors for gene therapy; nucleic acid constructs and viral vectors for use in cell therapy; nucleic acid constructs and viral vectors for use in genome editing; nucleic acid constructs and viral vectors for use in therapy Apparatus and instruments for the manufacture and processing of nucleic acid constructs, namely, DNA amplification apparatus and instruments for scientific and research purposes; nucleic acid constructs comprising natural and unnatural nucleotides, namely, nucleic acid amplification apparatus for scientific purposes; nucleic acid constructs comprising synthetic backbones or a mixture of natural and synthetic backbones, namely, nucleic acid amplification apparatus for scientific purposes; composites of nucleic acid constructs with organic and inorganic chemicals, namely, nucleic acid amplification apparatus for scientific purposes; all the aforesaid goods for use in nanotechnology applications including data storage, data retrieval and computing; electronic components in the form of electrical wires, semi-conductors, transistors, rectifiers, diodes, light emitting diodes (LEDs), and systems for energy storage, namely, batteries, all containing nucleic acid constructs; apparatus that uses nucleic acid constructs for labeling, imaging, and sensing applications for scientific research purposes; nucleic acid constructs as smart materials, being components of devices used for labeling, imaging and sensing DNA sequences for scientific research purposes; replacement parts for all the aforesaid technology Treatment of nucleic acid constructs; manufacture for others of nucleic acid constructs; manufacture for others of nucleic acids; manufacture for others of viruses; manufacture for others of vectors; manufacture for others of medical and veterinary therapies, vaccines, prophylactics, and counteragents; manufacture for others of nucleic acid constructs for nanotechnology applications; manufacture for others of nucleic acid constructs for labeling, imaging, and sensing applications; manufacture for others of nucleic acid constructs for electronic components; consultancy relating to the custom manufacture of the aforementioned goods Scientific and technological services, namely, research and design in the fields of chemicals, enzymes, nucleic acid constructs, and viral vectors; chemical and biological analysis and research services; research into nucleic acids and nucleic acid constructs; research and development into the creation of nucleic acid constructs; research, analysis, design, and development relating to therapies, prophylactics, and counteragents based on nucleic acid constructs; research and development in the fields of gene therapy, diagnostic preparation, agribiotechnology, and nanotechnology; consultancy relating to the aforesaid services
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals, enzymes, nucleic acid constructs and viral vectors for scientific and research use; nucleic acid constructs and viral vectors for use in the manufacture of nucleic acids; nucleic acid constructs and viral vectors for manufacturing viruses; nucleic acid constructs and viral vectors for use in scientific research, agribiotechnology, and nanotechnology; all of the aforementioned goods not for medical or veterinary use Nucleic acid constructs for medical and veterinary purposes; nucleic acid constructs and viral vectors for use as prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for the production of prophylactic or therapeutic vaccines; nucleic acid constructs and viral vectors for medical use; nucleic acid constructs and viral vectors for the production of therapeutic or prophylactic viruses; nucleic acid constructs and viral vectors for gene therapy; nucleic acid constructs and viral vectors for use in cell therapy; nucleic acid constructs and viral vectors for use in genome editing; nucleic acid constructs and viral vectors for use in therapy Scientific and technological services, namely, research and design in the fields of chemicals, enzymes, nucleic acid constructs, and viral vectors; chemical and biological analysis and research services; research into nucleic acids and nucleic acid constructs; research and development into the creation of nucleic acid constructs; research, analysis, design, and development relating to therapies, prophylactics, and counteragents based on nucleic acid constructs; research and development in the fields of gene therapy, diagnostic preparation, agribiotechnology, and nanotechnology; consultancy relating to the aforesaid services
The present invention concerns compositions and methods for the in vivo modification of cells of the immune system, in particular immune effector cells and immune memory cells to achieve therapeutic or prophylactic objectives. In particular, the novel method provides for the recruitment and in vivo modification of immune cells with a nucleic acid. Preferably, this enables the immune cells to express an antigen receptor. The novel method allows for increased yield of viable in vivo genetically modified immune cells, notably immune effector cells and/or immune memory cells, compared to prior art methods. Such transfected immune cells may have utility in treatment or prophylaxis.
The present invention relates to improved processes for production of closed linear deoxyribonucleic acid (DNA), in particular cell-free enzymatic production of closed linear DNA molecules, preferable using a closed linear DNA as a template for DNA synthesis. The invention further relates to a novel closed linear DNA species, suitable for use as a template in the improved processes for production of closed linear DNA. Further, the invention pertains to the intermediate products of the processes, since this enables the production of larger quantities of closed linear DNA from the template than with methods known in the art.
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.
The present invention relates to novel nucleotide complexes which enhance the enzymatic production of DNA. The nucleotide complexes includes a divalent cation at a ratio of 0.2-1.5 divalent cations per nucleotide, whilst monovalent ions (other than hydrogen or hydronium ions from any solvent) are minimal. The nucleotide complexes have desirable properties for the synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale. Further, the invention includes improved processes for preparing said nucleotide complexes. Further complexes are disclosed which possess unique properties in accelerating DNA synthesis at higher concentrations, these complexes also containing zwitterion molecules.
C01F 1/00 - Procédés de préparation des composés des métaux béryllium, magnésium, aluminium, calcium, strontium, baryum, radium, thorium ou des terres rares, en général
The present invention relates to novel nucleotide complexes which enhance the enzymatic production of DNA. The nucleotide complexes includes a divalent cation at a ratio of 0.2-1.5 divalent cations per nucleotide, whilst monovalent ions (other than hydrogen or hydronium ions from any solvent) are minimal. The nucleotide complexes have desirable properties for the synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale. Further, the invention includes improved processes for preparing said nucleotide complexes. Further complexes are disclosed which possess unique properties in accelerating DNA synthesis at higher concentrations, these complexes also containing zwitterion molecules.
C07H 23/00 - Composés contenant du bore, du silicium ou un métal, p. ex. chélates ou vitamine B12
C07H 19/10 - Radicaux pyrimidine avec le radical saccharide estérifié par des acides phosphoriques ou polyphosphoriques
C07H 19/20 - Radicaux purine avec le radical saccharide estérifié par des acides phosphoriques ou polyphosphoriques
C01F 1/00 - Procédés de préparation des composés des métaux béryllium, magnésium, aluminium, calcium, strontium, baryum, radium, thorium ou des terres rares, en général
The present invention relates to a novel closed linear DNA vector, which is suitable for use in the production of lentiviral particles. Notably, the present invention relates to a new configuration of the vector including the transgene (often termed the "payload" vector), which enables a greater yield of infectious lentiviral particles, notably a greater yield of lentiviral particles carrying a transgene, to be prepared when compared to closed linear DNA vectors lacking this configuration. Further, the inventors have developed improvements in lentiviral production with closed linear DNA, through optimisation of vector input quantities and construct ratios. The invention furthermore relates to a method of generating infectious lentiviral particles using the construct, optionally in conjunction with improved production vectors and/or optimised methodology.
The present invention relates to a novel closed linear DNA vector, which is suitable for use in the production of lentiviral particles. Notably, the present invention relates to a new configuration of the vector including the transgene (often termed the "payload" vector), which enables a greater yield of infectious lentiviral particles, notably a greater yield of lentiviral particles carrying a transgene, to be prepared when compared to closed linear DNA vectors lacking this configuration. Further, the inventors have developed improvements in lentiviral production with closed linear DNA, through optimisation of vector input quantities and construct ratios. The invention furthermore relates to a method of generating infectious lentiviral particles using the construct, optionally in conjunction with improved production vectors and/or optimised methodology.
An in vitro process for the production of closed linear deoxyribonucleic acid (DNA) comprises (a) contacting a DNA template comprising at least one protelomerase target sequence with at least one DNA polymerase in the presence of one or more primers under conditions promoting amplification of the template; and (b) contacting amplified DNA produced in (a) with at least one protelomerase under conditions promoting production of closed linear DNA. A kit provides components necessary in the process.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
A61K 39/00 - Préparations médicinales contenant des antigènes ou des anticorps
A61K 48/00 - Préparations médicinales contenant du matériel génétique qui est introduit dans des cellules du corps vivant pour traiter des maladies génétiquesThérapie génique
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Scientific and technological services, namely, research and design in the field of scientific DNA research and development; chemical and biological analysis and research services; Scientific research into DNA; Scientific research and development into the creation of DNA extracts and constructs; Scientific research, analysis, design and development relating to DNA based drugs and therapies; Scientific research and development in the fields of gene therapy, diagnostics, agribiotechnology and nanotechnology; consultancy relating to DNA research for scientific purposes
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Scientific and technological services, namely, research and design in the field of scientific DNA research and development; chemical and biological analysis and research services; Scientific research into DNA; Scientific research and development into the creation of DNA extracts and constructs; Scientific research, analysis, design and development relating to DNA based drugs and therapies; Scientific research and development in the fields of gene therapy, diagnostics, agribiotechnology and nanotechnology; consultancy relating to DNA research for scientific purposes
The present invention relates to an in vitro cell-free process for production of deoxyribonucleotides (DNAs) comprising at least one hairpin, corresponding DNA products and uses thereof, and oligonucleotides and kits useful in the process of the invention.
The present invention concerns new nucleic acid molecules which may be used in many applications, and methods for making the same. These nucleic acid molecules are preferably DNA vectors, optionally DNA expression vectors. The nucleic acid molecules are able to target the vector to a specific cellular location, such as the nucleus, due to the presence of one or more particular binding motifs within the nucleic acid molecule itself. Thus, the nucleic acid molecules of the invention may also be described as targeted delivery vectors, notably self-targeted delivery vectors or smart delivery vectors.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
Chemical preparations, enzymes and DNA for the preparation
of nucleic acid for scientific research; chemicals for use
in the preparation and creation of DNA extracts and
cassettes for scientific and industrial purposes; enzymes
for use in the preparation and creation of DNA extracts and
cassettes for scientific and industrial purposes; DNA
extracts and cassettes for scientific and research use; DNA
extracts for use in agri-biotechnology and nanotechnology;
nucleic acids comprising natural and unnatural nucleotides
for use in nanotechnology applications including data
storage, data retrieval and computing; nucleic acid
analogues comprising synthetic backbones or a mixture of
natural and synthetic backbones for use in nanotechnology
applications including data storage, data retrieval and
computing; composites of nucleic acids or nucleic acid
analogues with organic and inorganic chemicals for use in
nanotechnology applications including data storage, data
retrieval and computing; nucleic acid analogues with organic
and inorganic chemicals for laboratory use in labelling,
imaging and sensing applications; nucleic acid analogues
with organic and inorganic chemicals for laboratory use in
smart materials; two-dimensional and three-dimensional
nucleic acid analogues with organic and inorganic chemicals
for laboratory use in creating nanostructures and scaffolds;
nucleic acids or nucleic acid analogues for labelling,
imaging and sensing applications for scientific and research
purposes; nucleic acids or nucleic acid analogues as smart
materials for scientific and research purposes;
two-dimensional and three-dimensional nucleic acids or
nucleic acid analogues for creating nanostructures and
scaffolds for scientific and research purposes; parts and
fittings for all the aforesaid technology; all the aforesaid
goods for use in nanotechnology applications including data
storage, data retrieval and computing. Vaccines; DNA based vaccines; DNA extracts, namely, nucleic
acid sequences and chemical reagents for medical use;
therapeutic DNA, namely, nucleic acid sequences and chemical
reagents for medical and veterinary purposes; nucleic acids
comprising natural and unnatural nucleotides for medical and
veterinary purposes; nucleic acid analogues comprising
synthetic backbones or a mixture of natural and synthetic
backbones for medical and veterinary purposes; composites of
nucleic acids or nucleic acid analogues with organic and
inorganic chemicals for medical and veterinary purposes;
nucleic acid analogues with organic and inorganic chemicals
for medical and veterinary purposes in labelling, imaging
and sensing applications; nucleic acid analogues with
organic and inorganic chemicals for medical and veterinary
purposes in smart materials; two-dimensional and
three-dimensional nucleic acid analogues with organic and
inorganic chemicals for medical and veterinary purposes in
creating nanostructures and scaffolds. Electronic components and applications including wires,
semi-conductors including transistors, rectifiers, diodes
including light emitting diodes and systems for energy
storage such as batteries all comprising of nucleic acids or
nucleic acid analogues; parts and fittings for all the
aforesaid technology.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
Chemical preparations, enzymes and DNA for the preparation
of nucleic acid for scientific research; chemicals for use
in the preparation and creation of DNA extracts and
cassettes for scientific and industrial purposes; enzymes
for use in the preparation and creation of DNA extracts and
cassettes for scientific and industrial purposes; DNA
extracts and cassettes for scientific and research use; DNA
extracts for use in agri-biotechnology and nanotechnology;
nucleic acids comprising natural and unnatural nucleotides
for use in nanotechnology applications including data
storage, data retrieval and computing; nucleic acid
analogues comprising synthetic backbones or a mixture of
natural and synthetic backbones for use in nanotechnology
applications including data storage, data retrieval and
computing; composites of nucleic acids or nucleic acid
analogues with organic and inorganic chemicals for use in
nanotechnology applications including data storage, data
retrieval and computing; nucleic acid analogues with organic
and inorganic chemicals for laboratory use in labelling,
imaging and sensing applications; nucleic acid analogues
with organic and inorganic chemicals for laboratory use in
smart materials; two-dimensional and three-dimensional
nucleic acid analogues with organic and inorganic chemicals
for laboratory use in creating nanostructures and scaffolds;
nucleic acids or nucleic acid analogues for labelling,
imaging and sensing applications for scientific and research
purposes; nucleic acids or nucleic acid analogues as smart
materials for scientific and research purposes;
two-dimensional and three-dimensional nucleic acids or
nucleic acid analogues for creating nanostructures and
scaffolds for scientific and research purposes; parts and
fittings for all the aforesaid technology. Vaccines; DNA based vaccines; DNA extracts, namely, nucleic
acid sequences and chemical reagents for medical use;
therapeutic DNA, namely, nucleic acid sequences and chemical
reagents for medical and veterinary purposes; nucleic acids
comprising natural and unnatural nucleotides for medical and
veterinary purposes; nucleic acid analogues comprising
synthetic backbones or a mixture of natural and synthetic
backbones for medical and veterinary purposes; composites of
nucleic acids or nucleic acid analogues with organic and
inorganic chemicals for medical and veterinary purposes;
nucleic acid analogues with organic and inorganic chemicals
for medical and veterinary purposes in labelling, imaging
and sensing applications; nucleic acid analogues with
organic and inorganic chemicals for medical and veterinary
purposes in smart materials; two-dimensional and
three-dimensional nucleic acid analogues with organic and
inorganic chemicals for medical and veterinary purposes in
creating nanostructures and scaffolds. Electronic components and applications including wires,
semi-conductors including transistors, rectifiers, diodes
including light emitting diodes and systems for energy
storage such as batteries all comprising of nucleic acids or
nucleic acid analogues; parts and fittings for all the
aforesaid technology.
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large scale, with an improved yield and/or with an improved efficiency. The species of cation present in the nucleotide salt as the counter-ion is critical to the yield, efficiency and fidelity of high yielding enzymatic DNA synthesis reaction. The processes herein use alternative cations as counter-ions for the ionic nucleotides, permitting the use of higher concentrations of nucleotides in DNA synthesis, and further allowing for more favourable reaction conditions to be used.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
Chemical preparations, enzymes and DNA for the preparation of nucleic acid for scientific research, namely nucleic acid constructs; chemicals for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; enzymes for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in agri-biotechnology and nanotechnology, namely nucleic acid constructs other than for medical and veterinary purposes; nucleic acids comprising natural and unnatural nucleotides for use in nanotechnology applications including data storage, data retrieval and computing, namely nucleic acid constructs other than for medical and veterinary purposes; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for use in nanotechnology applications including data storage, data retrieval and computing other than for medical and veterinary purposes; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals for use in nanotechnology applications including data storage, data retrieval and computing, namely nucleic acid constructs other than for medical and veterinary purposes; nucleic acid analogues with organic and inorganic chemicals for laboratory use in labelling, imaging and sensing applications; nucleic acid analogues with organic and inorganic chemicals for laboratory use in smart materials; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals for laboratory use in creating nanostructures and scaffolds; nucleic acids and nucleic acid analogues for labelling, imaging and sensing applications for scientific and research purposes; nucleic acids and nucleic acid analogues as smart materials for scientific and research purposes; two-dimensional and three-dimensional nucleic acids and nucleic acid analogues for creating nanostructures and scaffolds for scientific and research purposes; parts and fittings for all the aforesaid technology; all the aforesaid goods for use in nanotechnology applications including data storage, data retrieval and computing Vaccines; DNA based vaccines; DNA extracts, namely, nucleic acid sequences and chemical reagents for medical use; therapeutic DNA, namely, nucleic acid sequences and chemical reagents for medical and veterinary purposes; nucleic acids comprising natural and unnatural nucleotides for medical and veterinary purposes; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for medical and veterinary purposes; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes; nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in labelling, imaging and sensing applications; nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in smart materials; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in creating nanostructures and scaffolds Electronic components, namely, nano-wires, semi-conductors, transistors, rectifiers, diodes, and batteries all the aforesaid goods being manufactured from or including nucleic acids, nucleic acid constructs and nucleic acid analogues; structural parts and fittings for all the aforesaid goods
09 - Appareils et instruments scientifiques et électriques
Produits et services
Scientific apparatus and instruments; apparatus for amplifying DNA; apparatus for manufacturing DNA; apparatus for processing DNA; parts and fittings for all the aforesaid goods
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.
Reprogramming allows the “conversion” of any mature or somatic cell of the human or animal body into a pluripotent stem cell. Reprogramming can be performed through the introduction of exogenous factors, usually transcription factors, into the mature cell. This process allows the production of induced pluripotent stem cells without the use of embryos, with the advantage that they can be produced from an individual to return/re-implant to the same individual. The inventors have developed a method of transient expression of exogenous reprogramming factors using a transient vector, wherein the vector is a closed linear DNA. Surprisingly, pluripotent stem cells developed in this manner are stable and closer in phenotype to natural stem cells such as ESCs.
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large scale, with an improved yield and/or with an improved efficiency. The species of cation present in the nucleotide salt as the counter-ion is critical to the yield, efficiency and fidelity of high yielding enzymatic DNA synthesis reaction. The processes herein use alternative cations as counter-ions for the ionic nucleotides, permitting the use of higher concentrations of nucleotides in DNA synthesis, and further allowing for more favourable reaction conditions to be used.
The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large scale, with an improved yield and/or with an improved efficiency. The species of cation present in the nucleotide salt as the counter-ion is critical to the yield, efficiency and fidelity of high yielding enzymatic DNA synthesis reaction. The processes herein use alternative cations as counter-ions for the ionic nucleotides, permitting the use of higher concentrations of nucleotides in DNA synthesis, and further allowing for more favourable reaction conditions to be used.
The present invention relates to an improved process for synthesis of DNA, RNA, proteins and like molecules, in particular cell-free enzymatic synthesis of DNA, preferably in large scale. The present invention relates to the synthesis of DNA using strand-displacement replication and the addition of nucleotides to the reaction mixture is controlled, thus controlling yield. The reaction mixture contains a starting amount of nucleotides, polymerase and DNA template, to which further nucleotides are supplied in a controlled manner.
Reprogramming allows the "conversion" of any mature or somatic cell of the human or animal body into a pluripotent stem cell. Reprogramming can be performed through the introduction of exogenous factors, usually transcription factors, into the mature cell. This process allows the production of induced pluripotent stem cells without the use of embryos, with the advantage that they can be produced from an individual to return/re-implant to the same individual.The inventors have developed a method of transient expression of exogenous reprogramming factors using a transient vector, wherein the vector is a closed linear DNA. Surprisingly, pluripotent stem cells developed in this manner are stable and closer in phenotype to natural stem cells such as ESCs.
Reprogramming allows the "conversion" of any mature or somatic cell of the human or animal body into a pluripotent stem cell. Reprogramming can be performed through the introduction of exogenous factors, usually transcription factors, into the mature cell. This process allows the production of induced pluripotent stem cells without the use of embryos, with the advantage that they can be produced from an individual to return/re-implant to the same individual.The inventors have developed a method of transient expression of exogenous reprogramming factors using a transient vector, wherein the vector is a closed linear DNA. Surprisingly, pluripotent stem cells developed in this manner are stable and closer in phenotype to natural stem cells such as ESCs.
An in vitro process for the production of closed linear deoxyribonucleic acid (DNA) comprises (a) contacting a DNA template comprising at least one protelomerase target sequence with at least one DNA polymerase in the presence of one or more primers under conditions promoting amplification of the template; and (b) contacting amplified DNA produced in (a) with at least one protelomerase under conditions promoting production of closed linear DNA. A kit provides components necessary in the process.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12Q 1/6853 - Réactions d’amplification d’acides nucléiques utilisant des amorces ou des matrices modifiées
C12N 15/11 - Fragments d'ADN ou d'ARNLeurs formes modifiées
A61K 39/00 - Préparations médicinales contenant des antigènes ou des anticorps
A61K 48/00 - Préparations médicinales contenant du matériel génétique qui est introduit dans des cellules du corps vivant pour traiter des maladies génétiquesThérapie génique
C12N 9/02 - Oxydoréductases (1.), p. ex. luciférase
The present invention relates to improved processes for production of closed linear deoxyribonucleic acid (DNA), in particular cell-free enzymatic production of closed linear DNA molecules, preferably using a closed linear DNA as a template for DNA synthesis. The invention further relates to a novel closed linear DNA species, suitable for use as a template in the improved processes for production of closed linear DNA. Further, the invention pertains to the intermediate products of the processes, since this enables the production of larger quantities of closed linear DNA from the template than with methods known in the art.
C07H 21/00 - Composés contenant au moins deux unités mononucléotide comportant chacune des groupes phosphate ou polyphosphate distincts liés aux radicaux saccharide des groupes nucléoside, p. ex. acides nucléiques
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
Produits et services
Chemical preparations, enzymes and DNA for the preparation of nucleic acid for scientific research; chemicals for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; enzymes for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in agri-biotechnology and nanotechnology; nucleic acids comprising natural and unnatural nucleotides for use in nanotechnology applications including data storage, data retrieval and computing; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for use in nanotechnology applications including data storage, data retrieval and computing; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals for use in nanotechnology applications including data storage, data retrieval and computing; nucleic acid analogues with organic and inorganic chemicals for laboratory use in labeling, imaging and sensing applications; nucleic acid analogues with organic and inorganic chemicals for laboratory use in smart materials; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals for laboratory use in creating nanostructures and scaffolds. Vaccines; DNA based vaccines; DNA extracts, namely, nucleic acid sequences and chemical reagents for medical use; therapeutic DNA, namely, nucleic acid sequences and chemical reagents for medical and veterinary purposes; nucleic acids comprising natural and unnatural nucleotides for medical and veterinary purposes; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for medical and veterinary purposes; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes; nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in labeling, imaging and sensing applications; nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in smart materials; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals for medical and veterinary purposes in creating nanostructures and scaffolds.
The present invention relates to improved processes for production of closed linear deoxyribonucleic acid (DNA), in particular cell-free enzymatic production of closed linear DNA molecules, preferably using a closed linear DNA as a template for DNA synthesis. The invention further relates to a novel closed linear DNA species, suitable for use as a template in the improved processes for production of closed linear DNA. Further, the invention pertains to the intermediate products of the processes, since this enables the production of larger quantities of closed linear DNA from the template than with methods known in the art.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
The present invention relates to an in vitro cell-free process for production of deoxyribonucleotides (DNAs) comprising at least one hairpin, corresponding DNA products and uses thereof, and oligonucleotides and kits useful in the process of the invention.
09 - Appareils et instruments scientifiques et électriques
Produits et services
Nucleic acids comprising natural and unnatural nucleotides; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals (NACs); all the aforesaid goods for use in nanotechnology applications including data storage, data retrieval and computing; electronic components and applications including NAC wires, semi-conductors including transistors, rectifiers, diodes including light emitting diodes and systems for energy storage such as batteries; NACs for labeling, imaging and sensing applications; NACs as smart materials; two-dimensional and three-dimensional NACs for creating nanostructures and scaffolds; parts and fittings for all the aforesaid technology.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
Produits et services
Nucleic acids comprising natural and unnatural nucleotides for laboratory use; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for laboratory use; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals (NACs) for laboratory use; all the aforesaid goods for use in nanotechnology applications [ including data storage, data retrieval and computing; nucleic acid analogues with organic and inorganic chemicals (NACs) for laboratory use in labeling, imaging and sensing applications; nucleic acid analogues with organic and inorganic chemicals (NACs) for laboratory use in smart materials; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals (NACs) for laboratory use in creating nanostructures and scaffolds ] Nucleic acids comprising natural and unnatural nucleotides for medical [ and veterinary ] purposes; nucleic acid analogues comprising synthetic backbones or a mixture of natural and synthetic backbones for medical [ and veterinary ] purposes; composites of nucleic acids or nucleic acid analogues with organic and inorganic chemicals (NACs) for medical [ and veterinary ] purposes; all the aforesaid goods for use in nanotechnology applications [ including data storage, data retrieval and computing; nucleic acid analogues with organic and inorganic chemicals (NACs) for medical and veterinary purposes in labeling, imaging and sensing applications ] ; nucleic acid analogues with organic and inorganic chemicals (NACs) for medical and veterinary purposes in smart materials [ ; two-dimensional and three-dimensional nucleic acid analogues with organic and inorganic chemicals (NACs) for medical and veterinary purposes in creating nanostructures and scaffolds ] [ Electronic components, namely, electric NAC wires, electronic semi-conductors, transistors, rectifiers, diodes, light emitting diodes, and batteries, all the aforesaid goods being manufactured from or including nucleic acid analogues with organic or inorganic chemicals (NACs); parts and fittings for all the aforesaid technology ]
The present invention relates to an in vitro cell-free process for production of deoxyribonucleotides (DNAs) comprising at least one hairpin, corresponding DNA products and uses thereof, and oligonucleotides and kits useful in the process of the invention.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
The present invention relates to an improved process for synthesis of DNA, RNA, proteins and like molecules, in particular cell-free enzymatic synthesis of DNA, preferably in large scale. The present invention relates to the synthesis of DNA using strand-displacement replication and the addition of nucleotides to the reaction mixture is controlled, thus controlling yield. The reaction mixture contains a starting amount of nucleotides, polymerase and DNA template, to which further nucleotides are supplied in a controlled manner.
C12M 1/36 - Appareillage pour l'enzymologie ou la microbiologie comportant une commande sensible au temps ou aux conditions du milieu, p. ex. fermenteurs commandés automatiquement
C12N 15/10 - Procédés pour l'isolement, la préparation ou la purification d'ADN ou d'ARN
C12P 19/34 - Polynucléotides, p. ex. acides nucléiques, oligoribonucléotides
The present invention relates to an improved process for synthesis of DNA, RNA, proteins and like molecules, in particular cell-free enzymatic synthesis of DNA, preferably in large scale. The present invention relates to the synthesis of DNA using strand-displacement replication and the addition of nucleotides to the reaction mixture is controlled, thus controlling yield. The reaction mixture contains a starting amount of nucleotides, polymerase and DNA template, to which further nucleotides are supplied in a controlled manner.
B01L 3/00 - Récipients ou ustensiles pour laboratoires, p. ex. verrerie de laboratoireCompte-gouttes
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
Apparatus for biochemical synthesis having a reaction vessel, a temperature control device for the reaction vessel, a plurality of reservoirs for reaction components, and supply/withdrawal systems, e.g. reciprocating syringe pumps and switchable valves. Agitation of a reaction mixture is effected by withdrawing a part of the mixture from the reaction vessel and returning it thereto.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemical preparations, enzymes and DNA for the preparation of nucleic acid for scientific research; chemicals for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; enzymes for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in [ agri-biotechnology and ] nanotechnology Vaccines; DNA based vaccines; DNA extracts, namely, nucleic acid sequences and chemical reagents for medical use; Therapeutic DNA, namely, nucleic acid sequences and chemical reagents for medical [ and veterinary ] purposes [ Scientific and technological services, namely, research and design in the field of DNA research and development; chemical and biological analysis and research services;scientific research in the field of DNA; scientific research and development in the field of creation of DNA extracts; scientific research, analysis, design and development relating to DNA based pharmaceutical drugs and therapies; scientific research and development in the fields of gene therapy, diagnostics, agri-biotechnology and nanotechnology; consultancy in the fields of gene therapy, diagnostics, agri-biotechnolgy, nano technology and DNA analysis ]
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemical preparations, enzymes and DNA for the preparation of nucleic acid for scientific research; chemicals for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; enzymes for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in [ agri-biotechnology and ] nanotechnology Vaccines; DNA based vaccines; DNA extracts, namely, nucleic acid sequences and chemical reagents for medical use; Therapeutic DNA, namely, nucleic acid sequences and chemical reagents for medical [ and veterinary ] purposes [ Scientific and technological services, namely, research and design in the field of DNA research and development; chemical and biological analysis and research services; scientific research in the field of DNA; scientific research and development in the field of creation of DNA extracts; scientific research, analysis, design and development relating to DNA based pharmaceutical drugs and therapies; scientific research and development in the fields of gene therapy, diagnostics, agri-biotechnology and nanotechnology; consultancy in the fields of gene therapy, diagnostics, agri-biotechnolgy, nano technology and DNA analysis ]
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
40 - Traitement de matériaux; recyclage, purification de l'air et traitement de l'eau
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals for scientific purposes; chemicals for use in
research; chemicals, enzymes and DNA for research use;
chemicals for use in the preparation and creation of DNA
extracts and cassettes; enzymes for use in the preparation
and creation of DNA extracts and cassettes; DNA extracts and
cassettes for scientific and research use; DNA extracts for
use in agri-biotechnology and nanotechnology. Pharmaceutical and veterinary products; vaccines; DNA based
vaccines; DNA extracts for medical use; therapeutic DNA. Scientific apparatus and instruments; apparatus for
amplifying DNA; apparatus for manufacturing DNA; apparatus
for processing DNA; parts and fittings for all the aforesaid
goods. Treatment of DNA to form DNA extracts; production of
therapeutic DNA by the manipulation of DNA into DNA extracts
or cassettes. Scientific and technological services and research and
design relating thereto; chemical and biological analysis
and research services; research in DNA; research and
development into the creation of DNA extracts; research,
analysis, design and development relating to DNA based drugs
and therapies; research and development in the fields of
gene therapy, diagnostics, agri-biotechnology and
nanotechnology; consultancy relating to the aforesaid
services.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals for scientific purposes; chemicals for use in research; chemicals, enzymes and DNA for research use; chemicals for use in the preparation and creation of DNA extracts and cassettes; enzymes for use in the preparation and creation of DNA extracts and cassettes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in agribiotechnology and nanotechnology. Pharmaceutical and veterinary products; vaccines; DNA based vaccines; DNA extracts for medical use; therapeutic DNA. Scientific and technological services and research and design relating thereto; chemical and biological analysis and research services; research into DNA; research and development into the creation of DNA extracts; research, analysis, design and development relating to DNA based drugs and therapies; research and development in the fields of gene therapy, diagnostics, agribiotechnology and nanotechnology; consultancy relating to the aforesaid services.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals for scientific purposes; chemicals for use in research; chemicals, enzymes and DNA for research use; chemicals for use in the preparation and creation of DNA extracts and cassettes; enzymes for use in the preparation and creation of DNA extracts and cassettes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in agribiotechnology and nanotechnology. Pharmaceutical and veterinary products; vaccines; DNA based vaccines; DNA extracts for medical use; therapeutic DNA. Scientific and technological services and research and design relating thereto; chemical and biological analysis and research services; research into DNA; research and development into the creation of DNA extracts; research, analysis, design and development relating to DNA based drugs and therapies; research and development in the fields of gene therapy, diagnostics, agribiotechnology and nanotechnology; consultancy relating to the aforesaid services.
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
40 - Traitement de matériaux; recyclage, purification de l'air et traitement de l'eau
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemical preparations, enzymes and DNA for the preparation of nucleic acid for scientific research; chemicals for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; enzymes for use in the preparation and creation of DNA extracts and cassettes for scientific and industrial purposes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in [ agri-biotechnology and ] nanotechnology Vaccines; DNA based vaccines; DNA extracts, namely, nucleic acid sequences and chemical reagents for medical use; Therapeutic DNA, namely, nucleic acid sequences and chemical reagents for medical [ and veterinary ] purposes [ Apparatus for amplifying DNA, namely, reactors and bioreactors for synthesising, processing, manufacturing, replicating and amplifying nucleic acid; apparatus for manufacturing DNA, namely, reactors and bioreactors for synthesising, processing, manufacturing, replicating and amplifying nucleic acid; apparatus for processing DNA, namely, reactors and bioreactors for synthesising, processing, manufacturing, replicating and amplifying nucleic acid ] Treatment of DNA to form DNA extracts; production of therapeutic DNA by the manipulation of DNA into DNA extracts or cassettes Scientific and technological services, namely, research and design in the field of DNA research and development; chemical and biological analysis and research services; scientific research in the field of DNA; scientific research and development in the field of creation of DNA extracts; scientific research, analysis, design and development relating to DNA based pharmaceutical drugs and therapies; scientific research and development in the fields of gene therapy, [ diagnostics, agri-biotechnology and nanotechnology; consultancy in the fields of gene therapy, diagnostics, agri-biotechnolgy, nano technology and DNA analysis ]
50.
Production of closed linear DNA using a palindromic sequence
A primer for the amplification of a DNA template comprising a protelomerase target sequence, particularly for production of closed linear DNA, which primer is capable of specifically binding to a palindromic sequence within a protelomerase target sequence and priming amplification in both directions.
C12Q 1/68 - Procédés de mesure ou de test faisant intervenir des enzymes, des acides nucléiques ou des micro-organismesCompositions à cet effetProcédés pour préparer ces compositions faisant intervenir des acides nucléiques
C12P 19/34 - Polynucléotides, p. ex. acides nucléiques, oligoribonucléotides
01 - Produits chimiques destinés à l'industrie, aux sciences ainsi qu'à l'agriculture
05 - Produits pharmaceutiques, vétérinaires et hygièniques
09 - Appareils et instruments scientifiques et électriques
40 - Traitement de matériaux; recyclage, purification de l'air et traitement de l'eau
42 - Services scientifiques, technologiques et industriels, recherche et conception
Produits et services
Chemicals for scientific purposes; chemicals for use in research; chemicals, enzymes and DNA for research use; chemicals for use in the preparation and creation of DNA extracts and cassettes; enzymes for use in the preparation and creation of DNA extracts and cassettes; DNA extracts and cassettes for scientific and research use; DNA extracts for use in agribiotechnology and nanotechnology. Pharmaceutical and veterinary products; vaccines; DNA based vaccines; DNA extracts for medical use; therapeutic DNA. Scientific apparatus and instruments; apparatus for amplifying DNA; apparatus for manufacturing DNA; apparatus for processing DNA; parts and fittings for all the aforesaid goods. Treatment of DNA to form DNA extracts; production of therapeutic DNA by the manipulation of DNA into DNA extracts or cassettes. Scientific and technological services and research and design relating thereto; chemical and biological analysis and research services; research into DNA; research and development into the creation of DNA extracts; research, analysis, design and development relating to DNA based drugs and therapies; research and development in the fields of gene therapy, diagnostics, agribiotechnology and nanotechnology; consultancy relating to the aforesaid services.
An in vitro process for the production of closed linear deoxyribonucleic acid (DNA) comprises (a) contacting aDNA template comprising at least one protelomerase target sequence with at least one DNA polymerase in the presence of one ormore primers under conditions promoting amplification of said template; and (b) contacting amplified DNA produced in (a) withat least one protelomerase under conditions promoting production of closed linear DNA. A kit provides components necessary inthe process.
