Plasmid-based one-pot saturation mutagenesis

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

doi:10.1038/nmeth.4029

Plasmid-based one-pot saturation mutagenesis

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

^*Corresponding author for this work

Chemical and Biological Engineering

Research output: Contribution to journal › Article › peer-review

111 Scopus citations

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.

Original language	English (US)
Pages (from-to)	928-930
Number of pages	3
Journal	Nature Methods
Volume	13
Issue number	11
DOIs	https://doi.org/10.1038/nmeth.4029
State	Published - Nov 1 2016

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1038/nmeth.4029

Cite this

@article{0223238c807340b99a7dba1963b096f6,

title = "Plasmid-based one-pot saturation mutagenesis",

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.",

author = "Wrenbeck, {Emily E.} and Klesmith, {Justin R.} and Stapleton, {James A.} and Adebola Adeniran and Tyo, {Keith E.J.} and Whitehead, {Timothy A.}",

year = "2016",

month = nov,

day = "1",

doi = "10.1038/nmeth.4029",

language = "English (US)",

volume = "13",

pages = "928--930",

journal = "Nature Methods",

issn = "1548-7091",

publisher = "Nature Research",

number = "11",

}

TY - JOUR

T1 - Plasmid-based one-pot saturation mutagenesis

AU - Wrenbeck, Emily E.

AU - Klesmith, Justin R.

AU - Stapleton, James A.

AU - Adeniran, Adebola

AU - Tyo, Keith E.J.

AU - Whitehead, Timothy A.

PY - 2016/11/1

Y1 - 2016/11/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84990878463&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84990878463&partnerID=8YFLogxK

U2 - 10.1038/nmeth.4029

DO - 10.1038/nmeth.4029

M3 - Article

C2 - 27723752

AN - SCOPUS:84990878463

SN - 1548-7091

VL - 13

SP - 928

EP - 930

JO - Nature Methods

JF - Nature Methods

IS - 11

ER -

Plasmid-based one-pot saturation mutagenesis

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this