Methods and compositions for cloning nucleic acid molecules

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435 911, 435 912, 435 915, 435 9152, 435184, 435194, 435471, C12N 1564, C12N 1574, C12N 999, C12N 912, C12P 1934

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061400866

ABSTRACT:
The present invention is directed generally to methods facilitating the cloning of nucleic acid molecules. In particular, the invention relates to the use of polymerase inhibitors, including but not limited to anti-polymerase antibodies (such as anti-Taq antibodies) and fragments thereof, to inactivate residual polymerase activity remaining after the amplification (particularly via PCR) of a target nucleic acid molecule. The invention further provides compositions, particularly storage-stable compositions, comprising one or more components, such as one or more restriction endonucleases and one or more polymerase inhibitors, that are useful in cloning amplified or synthesized nucleic acid molecules by the above-described methods. The invention also relates to nucleic acid molecules produced by these methods, and to genetic constructs (such as vectors) and host cells comprising these nucleic acid molecules.

REFERENCES:
patent: 5338671 (1994-08-01), Scalice et al.
patent: 5436149 (1995-07-01), Barnes
patent: 5512462 (1996-04-01), Cheng
patent: 5587287 (1996-12-01), Scalice et al.
patent: 5814502 (1998-09-01), Hoeltke et al.
Boehringer Mannheim Reagents for Molecular Biology catalog, 1990, pp. 15, 23, 1990.
Buchman, G.W. et al., "Rapid and Efficient Cloning of PCR Products Using the CloneAmp.TM.System," Focus 14:41-45 (1992).
Kaufman, D.L. and G.A. Evans, "Restriction Endonuclease Cleavage at the Termini of PCR Products," BioTechniques 9:304, 306 (19900.
Mead, D.A. et al., "A Universal Method for the Direct Cloning of PCR Amplified Nucleic Acid," Bio/Technology 9:657-663 (1991).
Polayes, D. and A.J. Hughes, Jr., "Efficient Protein Expression and Simple Purification Using the pProEX-1.TM.System," Focus 16:81-84 (1994).
Scharf, S.J., "Cloning with PCR, " in PCR Protocols: A Guide to Methods and Applications, New York: Academic Press, Inc., pp. 84-91 (1990).
Wybranietz, W.A. and U. Lauer, "Distinct Combination of Purification Methods Dramatically Improves Cohesive-End Subcloning of PCR Products," BioTechniques 24:578-580 (Apr. 1998).
Barnes, W.M., "The fidelity of Taq polymerase catalyzing PCR is improved by an N-terminal deletion," Gene 112:29-35 (1992).
Dang, C., and Jayasena, S.D., "Oligonucleotide Inhibitors of Taq DNA Polymerase Facilitate Detection of Low Copy Number Targets by PCR," J. Mol. Biol. 264:268-278 (1996).
Flaman, J.--M., et al., "A rapid PCR fidelity assay," Nucl. Acids Res. 22:3259-3260 (1994).
Kainz, P., et al., "Specificity-Enhanced Hot-Start PCR: Addition of Double-Stranded DNA Fragments Adapted to the Annealing Temperature," BioTechniques 28:278-282 (2000).
Kellogg, D.E., et al., TaqStart Antibody.TM.: "Hot Start" PCR Facilitated by a Neutralizing Monoclonal Antibody Directed Against Taq DNA Polymerase, BioTechniques 16:1134-1137 (1994).
Lawyer, F.C., et al., "High-level Expression, Purification, and Enzymatic Characterization of Full-Length Thermus aquaticus DNA Polymerase and a Truncated Form Deficient in 5'to 3'Exonuclease Activity," PCR Meth. Appl. 2:275-287 (1993).
Lin, Y., and Jayasena, S.D., "Inhibition of Multiple Thermostable DNA Polymerases by a Heterodimeric Aptamer," J. Mol. Biol. 271:100-111 (Aug. 1997).
New England Biolabs, Beverly, MA, product information for VENT.RTM., DEEPVENT.TM., and other thermophilic DNA polymerases, NEB 1996-1997 Products Catalogue, pp. 70-71 (1996).
Newton, C.R., "Thermostable DNA Polymerases," in: PCR Essential Data, C.R. Newton, Ed., New York: John Wiley & Sons, pp. 37-48 (1995).
Kahl, G., "Dictionary of Gene Technology," New York: VCH Publishers, Inc., pp. 140, 411, 510 (1995).
Life Technologies, Inc., "GIBCO BRL Products and Reference Guide 1997/1998," pp. 19-18 to 19-19 (1997).
Gerald, G.F., "Reverse Transcriptase: A Historical Perspective," Focus 20(3):65-67 (Fall 1998).
Bennett, B.L., and Molenaar, A.J., "Cloning of PCR Products Can be Inhibited by Taq Polymerase Carryover," BioTechniques 16:32,37 (Jan. 1994).
Fox, D., et al., "Prevention of Taq DNA Polymerase-Mediated Artifacts in PCR Product Cloning," Focus 20:15-16 (Mar. 1998).
Innis, M.A., et al., "DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA," Proc. Natl. Acad. Sci. USA 85:9436-9440 (Dec. 1988).
Nilsson, J., et al., "Heat-Mediated Activation of Affinity-Immobilized Taq DNA Polymerase," BioTechniques 22:744-751 (Apr. 1997).

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