TY - CHAP
T1 - Isolation of wheat genomic DNA for gene mapping and cloning
AU - Yu, Guotai
AU - Hatta, Asyraf
AU - Periyannan, Sambasivam
AU - Lagudah, Evans
AU - Wulff, Brande B.H.
N1 - Generated from Scopus record by KAUST IRTS on 2023-02-20
PY - 2017/1/1
Y1 - 2017/1/1
N2 - 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+).
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+).
UR - http://link.springer.com/10.1007/978-1-4939-7249-4_18
UR - http://www.scopus.com/inward/record.url?scp=85028988538&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7249-4_18
DO - 10.1007/978-1-4939-7249-4_18
M3 - Chapter
SP - 207
EP - 213
BT - Methods in Molecular Biology
PB - Humana Press [email protected]
ER -