河北农业大学学报
河北農業大學學報
하북농업대학학보
JOURNAL OF AGRICULTURAL UNIVERSITY OF HEBEI
2010年
2期
52-56
,共5页
红酵母%核酸提取%PCR扩增%RT-PCR
紅酵母%覈痠提取%PCR擴增%RT-PCR
홍효모%핵산제취%PCR확증%RT-PCR
Rhodotorula sp. RT-1%nucleic acid extraction%polymerase chain reaction%RT-PCR
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crtE cDNA内部序列,表明提取的RNA能够用于后续的分子生物学研究.
對蝸牛酶法、石英砂法、氯化芐法、SDS-CTAB法、反複凍融法、溶壁酶法等6種酵母DNA提取方法進行比較,確定瞭反複凍融法最適閤紅酵母RT-1的提取,所提取的DNA質量較高,以它為模闆成功剋隆到crtYB內部序列,證明其完全可用于酶切、測序等後續試驗.同時,比較瞭熱痠性酚法、酵母總RNA提取試劑盒 RNAsimple Total RNA Kit、硅藻土-苯酚法等3種酵母RNA提取方法,比較結果錶明:硅藻土-苯酚法所提RNA完整性高,用其進行RT-PCR剋隆得到crtE cDNA內部序列,錶明提取的RNA能夠用于後續的分子生物學研究.
대와우매법、석영사법、록화변법、SDS-CTAB법、반복동융법、용벽매법등6충효모DNA제취방법진행비교,학정료반복동융법최괄합홍효모RT-1적제취,소제취적DNA질량교고,이타위모판성공극륭도crtYB내부서렬,증명기완전가용우매절、측서등후속시험.동시,비교료열산성분법、효모총RNA제취시제합 RNAsimple Total RNA Kit、규조토-분분법등3충효모RNA제취방법,비교결과표명:규조토-분분법소제RNA완정성고,용기진행RT-PCR극륭득도crtE cDNA내부서렬,표명제취적RNA능구용우후속적분자생물학연구.
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.
