A Sequence- and Ligation-Independent Cloning (SLIC) procedure for the insertion of genes into a plasmid vector

Robert A. Holland

doi:10.1007/978-1-0716-3004-4_2

A Sequence- and Ligation-Independent Cloning (SLIC) procedure for the insertion of genes into a plasmid vector

Robert A. Holland

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

Abstract

Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase’s 3′ exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).

Original language	English
Title of host publication	DNA Manipulation and Analysis
Editors	Garry Scarlett
Place of Publication	New York
Publisher	Humana Press
Chapter	2
Pages	25-32
Number of pages	8
Edition	1st
ISBN (Electronic)	9781071630044
ISBN (Print)	9781071630037
DOIs	https://doi.org/10.1007/978-1-0716-3004-4_2
Publication status	Published - 1 Mar 2023

Publication series

Name	Methods in Molecular Biology
Volume	2633
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

SLIC
PCR
cloning
T4 polymerase
genetic engineering
protein expression

Access to Document

10.1007/978-1-0716-3004-4_2

Cite this

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abstract = "Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase{\textquoteright}s 3′ exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).",

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A Sequence- and Ligation-Independent Cloning (SLIC) procedure for the insertion of genes into a plasmid vector. / Holland, Robert A.
DNA Manipulation and Analysis. ed. / Garry Scarlett. 1st. ed. New York: Humana Press, 2023. p. 25-32 (Methods in Molecular Biology; Vol. 2633).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

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N2 - Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase’s 3′ exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).

AB - Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase’s 3′ exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).

KW - SLIC

KW - PCR

KW - cloning

KW - T4 polymerase

KW - genetic engineering

KW - protein expression

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