New rapid methods for DNA sequencing based in exonuclease III digestion followed by repair synthesis - PubMed
- ️Fri Jan 01 1982
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
-
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
-
Li C, Tucker PW. Li C, et al. Nucleic Acids Res. 1993 Mar 11;21(5):1239-44. doi: 10.1093/nar/21.5.1239. Nucleic Acids Res. 1993. PMID: 8464708 Free PMC article.
-
Site-specific nicking within the adenovirus inverted terminal repetition.
Chow KC, Pearson GD. Chow KC, et al. Nucleic Acids Res. 1984 Feb 10;12(3):1489-500. doi: 10.1093/nar/12.3.1489. Nucleic Acids Res. 1984. PMID: 6322107 Free PMC article.
-
Adenovirus origin of DNA replication: sequence requirements for replication in vitro.
Wides RJ, Challberg MD, Rawlins DR, Kelly TJ. Wides RJ, et al. Mol Cell Biol. 1987 Feb;7(2):864-74. doi: 10.1128/mcb.7.2.864-874.1987. Mol Cell Biol. 1987. PMID: 3821730 Free PMC article.
-
Domains of beta-tubulin essential for conserved functions in vivo.
Fridovich-Keil JL, Bond JF, Solomon F. Fridovich-Keil JL, et al. Mol Cell Biol. 1987 Oct;7(10):3792-8. doi: 10.1128/mcb.7.10.3792-3798.1987. Mol Cell Biol. 1987. PMID: 2891028 Free PMC article.
-
Exonuclease III protection assay with FRET probe for detecting DNA-binding proteins.
Wang J, Li T, Guo X, Lu Z. Wang J, et al. Nucleic Acids Res. 2005 Feb 1;33(2):e23. doi: 10.1093/nar/gni021. Nucleic Acids Res. 2005. PMID: 15687381 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases