Insertion and deletion mutagenesis by overlap extension PCR

Jehan Lee, Myeong Kyun Shin, Dong Kyun Ryu, Seahee Kim, Wang Shick Ryu

Research output: Chapter in Book/Report/Conference proceedingChapter

41 Citations (Scopus)

Abstract

Mutagenesis by the overlap extension PCR has become a standard method of creating mutations including substitutions, insertions, and deletions. Nonetheless, the established overlap PCR mutagenesis is limited in many respects. In particular, it has been difficult to make an insertion larger than 30 nt, since all sequence alterations must be embedded within the primer. Here, we describe a rapid and efficient method for creating insertions or deletions of any length at any position in a DNA molecule. This method is generally applicable, and therefore represents a significant improvement to the now widely used overlap extension PCR method.

Original languageEnglish
Title of host publicationIn Vitro Mutagenesis Protocols
Subtitle of host publicationThird Edition
EditorsBraman Jeff
Pages137-146
Number of pages10
DOIs
Publication statusPublished - 2010 Dec 1

Publication series

NameMethods in Molecular Biology
Volume634
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Lee, J., Shin, M. K., Ryu, D. K., Kim, S., & Ryu, W. S. (2010). Insertion and deletion mutagenesis by overlap extension PCR. In B. Jeff (Ed.), In Vitro Mutagenesis Protocols: Third Edition (pp. 137-146). (Methods in Molecular Biology; Vol. 634). https://doi.org/10.1007/978-1-60761-652-8_10