New rapid methods for DNA sequencing based in exonuclease III digestion followed by repair synthesis - PubMed

New rapid methods for DNA sequencing based in exonuclease III digestion followed by repair synthesis

L H Guo et al. Nucleic Acids Res. 1982.

Abstract

We describe improve enzymatic methods for sequencing method for sequencing DNA. They are based on partial digestion of duplex DNA with exonuclease III to produce DNA molecules with 3' ends shortened to varying lengths, followed by repair synthesis to extend and label the 3' ends. After asymmetrical cleavage of the DNA with a restriction enzyme, the labeled products are separated by gel electrophoresis and the sequence read from the autoradiogram. The entire procedures, beginning with unrestricted DNA and followed through gel electrophoresis, takes only one day for sequencing both strands of the DNA molecule. These methods are especially suitable for sequencing DNA cloned in plasmid vectors, and they greatly extend the usefulness of the dideoxynucleotide chain termination method of Sanger et al. (Proc. Natl. Acad. Sci. USA 74, 5463, 1977). Using these methods we have determined the sequence of a 410 base pair fragment which includes the yeast SUP3 tyrosine tRNA gene.

Similar articles

• Synchronous digestion of SV40 DNA by exonuclease III.

  Wu R, Ruben G, Siegel B, Jay E, Spielman P, Tu CP. Wu R, et al. Biochemistry. 1976 Feb 24;15(4):734-40. doi: 10.1021/bi00649a003. Biochemistry. 1976. PMID: 174717

• A DNA fragment with an alpha-phosphorothioate nucleotide at one end is asymmetrically blocked from digestion by exonuclease III and can be replicated in vivo.

  Putney SD, Benkovic SJ, Schimmel PR. Putney SD, et al. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7350-4. doi: 10.1073/pnas.78.12.7350. Proc Natl Acad Sci U S A. 1981. PMID: 6278470 Free PMC article.

• Exonuclease III of Escherichia coli K-12, an AP endonuclease.

  Rogers SG, Weiss B. Rogers SG, et al. Methods Enzymol. 1980;65(1):201-11. doi: 10.1016/s0076-6879(80)65028-9. Methods Enzymol. 1980. PMID: 6246343 No abstract available.

• [Enzymology of DNA repair processes].

  Tomilin NV. Tomilin NV. Tsitologiia. 1977 Oct;19(10):1086-1109. Tsitologiia. 1977. PMID: 341447 Review. Russian. No abstract available.

• Recent studies of the fidelity of DNA synthesis.

  Kunkel TA, Bebenek K. Kunkel TA, et al. Biochim Biophys Acta. 1988 Nov 10;951(1):1-15. doi: 10.1016/0167-4781(88)90020-6. Biochim Biophys Acta. 1988. PMID: 2847793 Review. No abstract available.

Cited by

• Fusion of adenovirus E1A to the glucocorticoid receptor by high-resolution deletion cloning creates a hormonally inducible viral transactivator.

  Becker DM, Hollenberg SM, Ricciardi RP. Becker DM, et al. Mol Cell Biol. 1989 Sep;9(9):3878-87. doi: 10.1128/mcb.9.9.3878-3887.1989. Mol Cell Biol. 1989. PMID: 2550806 Free PMC article.

• Systematic Probing of the Sequence Selectivity of Exonuclease III with a Photosensitization Colorimetric Assay.

  Ding Y, Li X, Zhang X, Li F, Hou X, Wu P. Ding Y, et al. ACS Omega. 2019 Aug 6;4(8):13382-13387. doi: 10.1021/acsomega.9b01560. eCollection 2019 Aug 20. ACS Omega. 2019. PMID: 31460466 Free PMC article.

• A poliovirus type 1 neutralization epitope is located within amino acid residues 93 to 104 of viral capsid polypeptide VP1.

  Wychowski C, van der Werf S, Siffert O, Crainic R, Bruneau P, Girard M. Wychowski C, et al. EMBO J. 1983;2(11):2019-24. doi: 10.1002/j.1460-2075.1983.tb01694.x. EMBO J. 1983. PMID: 6196195 Free PMC article.

• Isolation and characterization of two alleles of the chicken cytochrome c gene.

  Limbach KJ, Wu R. Limbach KJ, et al. Nucleic Acids Res. 1983 Dec 20;11(24):8931-50. doi: 10.1093/nar/11.24.8931. Nucleic Acids Res. 1983. PMID: 6324108 Free PMC article.

• Regulation, linkage, and sequence of mouse metallothionein I and II genes.

  Searle PF, Davison BL, Stuart GW, Wilkie TM, Norstedt G, Palmiter RD. Searle PF, et al. Mol Cell Biol. 1984 Jul;4(7):1221-30. doi: 10.1128/mcb.4.7.1221-1230.1984. Mol Cell Biol. 1984. PMID: 6095054 Free PMC article.

References

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Europe PubMed Central
  • PubMed Central
  • Silverchair Information Systems

• Other Literature Sources

  • The Lens - Patent Citations Database

• Molecular Biology Databases

  • Saccharomyces Genome Database