A partial-complementary adapter for an improved and simplified ligation-mediated suppression PCR technique

Ji U. Jeung, Ki Cho Sung, Jeong Sheop Shin

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Ligation-mediated suppression PCR (LMS-PCR) is a powerful tool for walking in unknown genomic DNA regions from known adjacent sequences. This approach has made it feasible to obtain promoter sequences and to enable researchers to identify full-length gene sequences or isoforms of multigene families. However, the advantages of LMS-PCR can be obviated by the presence of incomplete base modifications on the suppression adapters. We propose here that a 'partial-complementary adapter' is a more reliable suppression adapter, demanding only 5′-end phosphorylation. We also describe a simplified procedure for the easier preparation of PCR templates with very small quantities of DNA and a fast and direct characterization of the suppression-PCR products. A set of practical guidelines is proposed for pre-checking the efficiency of the adapter modification using two model systems: bacteriophage λ (λ) and Arabidopsis.

Original languageEnglish
Pages (from-to)110-120
Number of pages11
JournalJournal of Biochemical and Biophysical Methods
Volume64
Issue number2
DOIs
Publication statusPublished - 2005 Aug 31

Keywords

  • Chromosome walking
  • Ligation-mediated suppression PCR
  • Partial-complementary adapter

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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