摘要
对蜗牛酶法、石英砂法、氯化苄法、SDS-CTAB法、反复冻融法、溶壁酶法等6种酵母DNA提取方法进行比较,确定了反复冻融法最适合红酵母RT-1的提取,所提取的DNA质量较高,以它为模板成功克隆到crtYB内部序列,证明其完全可用于酶切、测序等后续试验。同时,比较了热酸性酚法、酵母总RNA提取试剂盒RNAsimple Total RNA Kit、硅藻土-苯酚法等3种酵母RNA提取方法,比较结果表明:硅藻土-苯酚法所提RNA完整性高,用其进行RT-PCR克隆得到crtEcDNA内部序列,表明提取的RNA能够用于后续的分子生物学研究。
Six methods of Rhodotorula sp.RT-1 genomic DNA extraction were compared,including snailase,quartz sand,benzyl chioride,SDS-CTAB,repeated freezing and lyticase methods.The results showed that repeated freezing was the most suitable method for extracting genomic DNA.As the PCR template,it has successfully amplified the ajm fragment in the gene of crtYB.At the same time,the methods of hot acidic phenol,RNAsimple Total RNA Kit and macaloid-phenol for RNA extraction were compared and it was found that the method of macaloid-phenol could get a higher Integrity of RNA.As the interior sequence of the crtE was amplified,the RNA gained was proved to be useful for future molecular biological research.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2010年第2期52-56,共5页
Journal of Hebei Agricultural University
基金
基础微生物菌种资源标准化、整理、整合与分享(2005DKA2120605)
河北省微生物资源基础信息库09967119D-16
关键词
红酵母
核酸提取
PCR扩增
RT-PCR
Rhodotorula sp.RT-1
nucleic acid extraction
polymerase chain reaction
RT-PCR
