[An efficient method for isolation of mitochondrial DNA in wheat].
An efficient method for isolation of mitochondrial DNA (mtDNA) from etiolated tissues of wheat was developed. The protocol consists of mitochondria isolation with differential centrifugation, Dnase I treatment, lysis with SDS and proteinase K, removing protein by TE-saturated phenol/chloroform extraction and a final RNase A treatment for obtaining mtDNA. The mtDNA samples were tested using spectrophotometry and agarose gel electrophoresis. It was proved that the mtDNA isolated by this method not only have the high yield but also structural complete, and contains no impurities, such as nuclear DNA, RNA and protein. The result showed that this high quality mtDNA can be successfully used in PCR and other genetic studies. In addition, it was found that adjusting the lysis temperature has a noticeable effect on the mtDNA yield.
Li
Wen-Qiang
WQ
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Education Ministry, Huazhong Agricultural University, Wuhan 430070, China. lwq54@163.com
Zhang
Gai-Sheng
GS
Wang
Kui
K
Niu
Na
N
Pan
Dong-Liang
DL
D004272:DNA, Mitochondrial / Q000302:isolation & purification*; D018744:DNA, Plant / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D019105:Random Amplified Polymorphic DNA Technique; D013053:Spectrophotometry; D014908:Triticum / Q000235:genetics*
D004272:DNA, Mitochondrial; D018744:DNA, Plant