Gene amplification and qRT-PCR

Cerith Jones, Alain Filloux

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

This chapter includes methods for the use of the polymerase chain reaction (PCR) with Pseudomonas, and several specific tips for their successful application in this organism. The first part of the chapter includes methods for purifying genomic DNA from, and amplifying genes from, Pseudomonas, in addition to methods which describe how to prepare a cell lysate from Pseudomonas species for colony PCR reactions. The chapter continues with a switch in focus from DNA to RNA, describing methods for RNA isolation from Pseudomonas, cDNA generation, and finally q-RT-PCR to investigate relative changes in gene expression.

Original languageEnglish
Title of host publicationPseudomonas Methods and Protocols
EditorsAlain Filloux, Juan-Luis Ramos
Pages457-468
Number of pages12
Volume1149
ISBN (Electronic)978-1-4939-0473-0
DOIs
Publication statusPublished - 2014

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1149
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CDNA
  • Cloning
  • Colony PCR
  • Gene amplification
  • PCR
  • QRT-PCR
  • RNA
  • RT-PCR

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