Plasmid-based one-pot saturation mutagenesis

Emily E. Wrenbeck, Justin R. Klesmith, James A. Stapleton, Adebola Adeniran, Keith E.J. Tyo, Timothy A. Whitehead

Research output: Contribution to journalArticle

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Abstract

Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped plasmid dsDNA. The method can be used to produce comprehensive or single- or multi-site saturation mutagenesis libraries.

LanguageEnglish (US)
Pages928-930
Number of pages3
JournalNature Methods
Volume13
Issue number11
DOIs
StatePublished - Nov 1 2016

Profile

Mutagenesis
Plasmids
Libraries
Scanning

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology

Cite this

Wrenbeck, E. E., Klesmith, J. R., Stapleton, J. A., Adeniran, A., Tyo, K. E. J., & Whitehead, T. A. (2016). Plasmid-based one-pot saturation mutagenesis. Nature Methods, 13(11), 928-930. DOI: 10.1038/nmeth.4029

Plasmid-based one-pot saturation mutagenesis. / Wrenbeck, Emily E.; Klesmith, Justin R.; Stapleton, James A.; Adeniran, Adebola; Tyo, Keith E.J.; Whitehead, Timothy A.

In: Nature Methods, Vol. 13, No. 11, 01.11.2016, p. 928-930.

Research output: Contribution to journalArticle

Wrenbeck, EE, Klesmith, JR, Stapleton, JA, Adeniran, A, Tyo, KEJ & Whitehead, TA 2016, 'Plasmid-based one-pot saturation mutagenesis' Nature Methods, vol 13, no. 11, pp. 928-930. DOI: 10.1038/nmeth.4029
Wrenbeck EE, Klesmith JR, Stapleton JA, Adeniran A, Tyo KEJ, Whitehead TA. Plasmid-based one-pot saturation mutagenesis. Nature Methods. 2016 Nov 1;13(11):928-930. Available from, DOI: 10.1038/nmeth.4029
Wrenbeck, Emily E. ; Klesmith, Justin R. ; Stapleton, James A. ; Adeniran, Adebola ; Tyo, Keith E.J. ; Whitehead, Timothy A./ Plasmid-based one-pot saturation mutagenesis. In: Nature Methods. 2016 ; Vol. 13, No. 11. pp. 928-930
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