In vivo cloning of PCR products in E.coli

Jonathan D. Oliner, Kenneth W Kinzler, Bert Vogelstein

Research output: Contribution to journalArticle

Abstract

This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).

Original languageEnglish (US)
Pages (from-to)5192-5197
Number of pages6
JournalNucleic Acids Research
Volume21
Issue number22
StatePublished - Nov 11 1993

Fingerprint

Cloning
Escherichia coli
Escherichia Coli
Organism Cloning
Polymerase Chain Reaction
Homologous Recombination
DNA
Clone
Recombination
Ligation
Clone Cells
Requirements
Therapeutics

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

In vivo cloning of PCR products in E.coli. / Oliner, Jonathan D.; Kinzler, Kenneth W; Vogelstein, Bert.

In: Nucleic Acids Research, Vol. 21, No. 22, 11.11.1993, p. 5192-5197.

Research output: Contribution to journalArticle

Oliner, Jonathan D. ; Kinzler, Kenneth W ; Vogelstein, Bert. / In vivo cloning of PCR products in E.coli. In: Nucleic Acids Research. 1993 ; Vol. 21, No. 22. pp. 5192-5197.
@article{caa5a78bb26444fab57c7b6095fa9125,
title = "In vivo cloning of PCR products in E.coli",
abstract = "This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).",
author = "Oliner, {Jonathan D.} and Kinzler, {Kenneth W} and Bert Vogelstein",
year = "1993",
month = "11",
day = "11",
language = "English (US)",
volume = "21",
pages = "5192--5197",
journal = "Nucleic Acids Research",
issn = "1362-4962",
publisher = "Oxford University Press",
number = "22",

}

TY - JOUR

T1 - In vivo cloning of PCR products in E.coli

AU - Oliner, Jonathan D.

AU - Kinzler, Kenneth W

AU - Vogelstein, Bert

PY - 1993/11/11

Y1 - 1993/11/11

N2 - This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).

AB - This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).

UR - http://www.scopus.com/inward/record.url?scp=0027385009&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027385009&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 5192

EP - 5197

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 1362-4962

IS - 22

ER -