Abstract:

In this experiment, high-salt differential centrifugation and differential centrifugation of sucrose sedimentating were used to extract mitochondrial DNA from carrot leaves. No nuclear DNA-contamined mitochondrial DNA was obtained by adding DNase Ⅰ and changing the force of centrifuging and sedimentating of mitochondrial. To crack mitochondria, SDS and proteinase K were used. Pure mitochondrial DNA was successfully obtained by phenol / chloroform / isoamyl alcohol extraction to remove protein, and digestion with RNase A.Chloroplast-specific primers were designed to detect mitochondrial DNA purity. The results showed that using the optimized method of differential centrifugation of sucrose sedimentating extracting carrot mitochondrial DNA not only operate simpler but also obtain mitochondrial DNA with higher purity and higher yield(34.46 μg mitochondrial DNA per gram of leaf ).