Isolation of wheat genomic DNA for gene mapping and cloning

Guotai Yu, Asyraf Hatta, Sambasivam Periyannan, Evans Lagudah, Brande B.H. Wulff

Research output: Chapter in Book/Report/Conference proceedingChapter

24 Scopus citations

Abstract

DNA is widely used in plant genetic and molecular biology studies. In this chapter, we describe how to extract DNA from wheat tissues. The tissue samples are ground to disrupt the cell wall. Then cetyltrimethylammonium bromide (CTAB) or sodium dodecyl sulfate (SDS) is used to disrupt the cell and nuclear membranes to release the DNA into solution. A reducing agent, β-mercaptoethanol, is added to break the disulfide bonds between the cysteine residues and to help remove the tanins and polyphenols. A high concentration of salt is employed to remove polysaccharides. Ethylenediaminetetraacetic acid (EDTA) stops DNase activity by chelating the magnesium ions. The nucleic acid solution is extracted with chloroform–isoamyl alcohol (24:1) or 6Mammonium acetate. The DNA in aqueous phase is precipated with ethanol or isopropanol, which makes DNA less hydrophilic in the presence of sodium ions (Na+).
Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.humana@humanapr.com
Pages207-213
Number of pages7
DOIs
StatePublished - Jan 1 2017
Externally publishedYes

Bibliographical note

Generated from Scopus record by KAUST IRTS on 2023-02-20

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Isolation of wheat genomic DNA for gene mapping and cloning'. Together they form a unique fingerprint.

Cite this