Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus

C. D. Herring, C. Chevillard, S. L. Johnston, P. J. Wettstein, R. Riblet

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We have developed a simple PCR strategy, termed vector-hexamer PCR, that is unique in its ability to easily recover every insert end from large insert clones in YAC and BAC vectors. We used this method to amplify and isolate all insert ends from a YAC contig covering the mouse Igh locus. Seventy-seven ends were amplified and sequenced from 36 YAC clones from four libraries in the pYAC4 vector. Unexpectedly, 40% of the insert ends of these YACs were LINE1 repeats. Nonrepetitive ends were suitable for use as probes on Southern blots of digested YACs to identify overlaps and construct a contig. The same strategy was used successfully to amplify insert ends from YACs in the pRML vector from the Whitehead Institute/MIT-820 mouse YAC library and from BACs in pBeloBAC11. The simplicity of this technique and its ability to isolate every end from large insert clones are of great utility in genomic investigation.

Original languageEnglish (US)
Pages (from-to)673-681
Number of pages9
JournalGenome Research
Volume8
Issue number6
StatePublished - 1998

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Clone Cells
Polymerase Chain Reaction
Libraries
Southern Blotting

ASJC Scopus subject areas

  • Genetics

Cite this

Herring, C. D., Chevillard, C., Johnston, S. L., Wettstein, P. J., & Riblet, R. (1998). Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus. Genome Research, 8(6), 673-681.

Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus. / Herring, C. D.; Chevillard, C.; Johnston, S. L.; Wettstein, P. J.; Riblet, R.

In: Genome Research, Vol. 8, No. 6, 1998, p. 673-681.

Research output: Contribution to journalArticle

Herring, CD, Chevillard, C, Johnston, SL, Wettstein, PJ & Riblet, R 1998, 'Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus', Genome Research, vol. 8, no. 6, pp. 673-681.
Herring CD, Chevillard C, Johnston SL, Wettstein PJ, Riblet R. Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus. Genome Research. 1998;8(6):673-681.
Herring, C. D. ; Chevillard, C. ; Johnston, S. L. ; Wettstein, P. J. ; Riblet, R. / Vector hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus. In: Genome Research. 1998 ; Vol. 8, No. 6. pp. 673-681.
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