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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