A DNA sequencing strategy - PubMed

A DNA sequencing strategy

Y M Lee et al. Anal Biochem. 1988 Dec.

Abstract

A modification of Lin's systematic DNA sequencing strategy is described. A method based on the religation of compatible cohesive ends generated by Sau3AI and BamHI was developed. The original procedure has been simplified and the yield of transfectant has been greatly improved. After complete digestion with BamHI and limited cleavage with Sau3AI, the single-cut linear DNA does not have to be separated from the supercoil or the open circular DNA on an agarose gel. After ligation, the DNA is digested with the restriction enzyme between the cloning site and BamHI site again. The original intact DNA is linearized, whereas the deleted subclone is not. Therefore the background is decreased to an undetectable level. This DNA sequencing strategy was tested on a 1.4-kb cDNA fragment containing the haptoglobin-related sequences. It is not necessary to purify large amounts of RF DNA (500 ng is enough) to get enough subclones. A set of subclones was produced in 1 day and the yield of plaques was about sixfold higher than that published.

Similar articles

• An improved DNA sequencing strategy.

  Lin HC, Lei SP, Wilcox G. Lin HC, et al. Anal Biochem. 1985 May 15;147(1):114-9. doi: 10.1016/0003-2697(85)90016-8. Anal Biochem. 1985. PMID: 2992313

• Generation of deletion subclones for sequencing by partial digestion with restriction endonucleases.

  Lamperti ED, Villa-Komaroff L. Lamperti ED, et al. Anal Biochem. 1990 Feb 15;185(1):187-93. doi: 10.1016/0003-2697(90)90278-h. Anal Biochem. 1990. PMID: 2160779

• Rapid method for identifying subclones bridging gaps between contiguous DNA sequences.

  Lu YY, Ni FD, Hong GF. Lu YY, et al. Chin J Biotechnol. 1989;5(3):157-60. Chin J Biotechnol. 1989. PMID: 2491324

• Efficient DNA subcloning through selective restriction endonuclease digestion.

  Spear MA. Spear MA. Biotechniques. 2000 Apr;28(4):660-2, 664, 666 passim. doi: 10.2144/00284st01. Biotechniques. 2000. PMID: 10769743

• A differential cloning procedure for rearranged or altered genomic DNA based on in-gel competitive reassociation.

  Kiyama R, Inoue S, Ohki R, Kikuya E, Yokota H, Oishi M. Kiyama R, et al. Adv Biophys. 1995;31:151-61. doi: 10.1016/0065-227x(95)99389-7. Adv Biophys. 1995. PMID: 7625271 Review.

Cited by

• Molecular cloning, nucleotide sequence and expression in Escherichia coli of the beta-cyclodextrin glycosyltransferase gene from Bacillus circulans strain no. 8.

  Nitschke L, Heeger K, Bender H, Schulz GE. Nitschke L, et al. Appl Microbiol Biotechnol. 1990 Aug;33(5):542-6. doi: 10.1007/BF00172548. Appl Microbiol Biotechnol. 1990. PMID: 1368573

• Nucleotide sequence of katF of Escherichia coli suggests KatF protein is a novel sigma transcription factor.

  Mulvey MR, Loewen PC. Mulvey MR, et al. Nucleic Acids Res. 1989 Dec 11;17(23):9979-91. doi: 10.1093/nar/17.23.9979. Nucleic Acids Res. 1989. PMID: 2690013 Free PMC article.

• Nucleotide and derived amino acid sequences of the major porin of Comamonas acidovorans and comparison of porin primary structures.

  Gerbl-Rieger S, Peters J, Kellermann J, Lottspeich F, Baumeister W. Gerbl-Rieger S, et al. J Bacteriol. 1991 Apr;173(7):2196-205. doi: 10.1128/jb.173.7.2196-2205.1991. J Bacteriol. 1991. PMID: 1848840 Free PMC article.

MeSH terms

Substances