Isolation of wheat genomic DNA for gene mapping and cloning

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

doi:10.1007/978-1-4939-7249-4_18

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 proceeding › Chapter

27 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 language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press [email protected]
Pages	207-213
Number of pages	7
DOIs	https://doi.org/10.1007/978-1-4939-7249-4_18
State	Published - Jan 1 2017
Externally published	Yes

Bibliographical note

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

ASJC Scopus subject areas

Genetics
Molecular Biology

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10.1007/978-1-4939-7249-4_18

Cite this

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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+).",

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AU - Yu, Guotai

AU - Hatta, Asyraf

AU - Periyannan, Sambasivam

AU - Lagudah, Evans

AU - Wulff, Brande B.H.

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AB - 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+).

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BT - Methods in Molecular Biology

PB - Humana Press [email protected]

ER -

Isolation of wheat genomic DNA for gene mapping and cloning

Abstract

Bibliographical note

ASJC Scopus subject areas

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