The method of DNA extraction from animal muscle tissue and real- time PCR detection

Genomic DNA of muscle tissues from fresh mutton,pork,chicken,beef,duck were extracted through SDS method,Gu SCN method and kit method.The DNA was detected by agarosegel electrophoresis and real- time PCR method.Results indicated that the kit method improved better concentration and purity of DNA than SDS method and Gu SCN method.By comparing the three extraction methods to establish standard curve,the sorted according to the quality of the regression coefficients: kit method≥Gu SCN method > SDS method.The amplification efficiency according to the sorting for quality was kit method ≥ Gu SCN method > SDS method. The detection sensitivity of sheep- derived ingredients sensitivity by three Extraction Methods,the minimum detection limits were 80 pg / μL,but in the low- concentration detecting the relative standard deviation( RSD) of the kit method 0.684% was less than Gu SCN method 0.734% and SDS method 1.075%.The detection of pig- derived ingredients sensitivity,the minimum detection limit of SDS method was 80 pg / μL.The minimum detection limit of Gu SCN method and kit method were16 pg / μL,but in the low- concentration detecting the relative standard deviation( RSD) of the kit method 0.092%was less than Gu SCN method 4.640%. The detection of chicken- derived ingredients sensitivity,the minimum detection limit of SDS method and Gu SCN method were 3.2 pg / μL,the minimum detection limit of kit method was640 fg / μL.Above all,the genomic DNA from kit method was more conducive to use the real- time PCR technique in food adulteration and species identification.