Comparison of methods for total microbial DNA extraction from solid-state fermentation
固态发酵过程中微生物总DNA提取方法比较

To analyze the diversity and succession of microbes during solid-state fermentation,different genomic DNA extraction methods for bacteria and fungi were employed,including lyticase lysis,ultrasonic lysis and grinding lysis in liquid nitrogen with CTAB.The quantity and purity of the DNA were measured by ultraviolet spectrophotometer.PCR amplification was carried out with a eubacterial 16SrDNA targeted primer pair(341F and 907R)and a fungi 18SrDNA targeted primer pair(NU-SSU-0817 and NU-SSU-1196).The bacterial and fungal diversity during solid-state fermentation was analyzed by denaturing gradient gel electrophoresis(DGGE).The results show that the crude and purified DNA fragments extracted during solid-state fermentation have a length of about 23 kb.The length of the DNA fragments after bacterial and fungal PCR amplification was about 586 bp and 422 bp,respectively.DNA band profiles by DGGE analysis for either bacterial or fungal PCR products testify to the similar microbial diversity observed for all three DNA extraction methods.Lyticase lysis yielded the highest quantity of DNA,followed by ultrasonic lysis,followed by grinding lysis in liquid nitrogen with CTAB.