湖北民族学院学报（自然科学版）
湖北民族學院學報（自然科學版）
호북민족학원학보（자연과학판）
JOURNAL OF HUBEI INSTITUTE FOR NATIONALITIES（NATURAL SCIENCE EDITION）
2015年
2期
170-173
,共4页
兼并引物%常规PCR%降落PCR%巢式PCR
兼併引物%常規PCR%降落PCR%巢式PCR
겸병인물%상규PCR%강락PCR%소식PCR
degenerate primer%conventional PCR%touchdown PCR%nested PCR
根据NCBI上的DNA拓扑异构酶Ⅱ基因序列设计兼并引物，采用常规PCR、降落PCR、巢式PCR三种方法扩增糙皮侧耳和阿魏侧耳的拓扑异构酶Ⅱ部分基因序列，比较三种方法扩增效果的特异性、灵敏度。结果表明，巢式PCR的灵敏度和特异性都优于常规PCR和降落PCR，并且巢式PCR的灵敏度是常规PCR灵敏度的100倍以上，更加适合兼并引物扩增。这对利用兼并引物获取特异基因具有指导意义。
根據NCBI上的DNA拓撲異構酶Ⅱ基因序列設計兼併引物，採用常規PCR、降落PCR、巢式PCR三種方法擴增糙皮側耳和阿魏側耳的拓撲異構酶Ⅱ部分基因序列，比較三種方法擴增效果的特異性、靈敏度。結果錶明，巢式PCR的靈敏度和特異性都優于常規PCR和降落PCR，併且巢式PCR的靈敏度是常規PCR靈敏度的100倍以上，更加適閤兼併引物擴增。這對利用兼併引物穫取特異基因具有指導意義。
근거NCBI상적DNA탁복이구매Ⅱ기인서렬설계겸병인물，채용상규PCR、강락PCR、소식PCR삼충방법확증조피측이화아위측이적탁복이구매Ⅱ부분기인서렬，비교삼충방법확증효과적특이성、령민도。결과표명，소식PCR적령민도화특이성도우우상규PCR화강락PCR，병차소식PCR적령민도시상규PCR령민도적100배이상，경가괄합겸병인물확증。저대이용겸병인물획취특이기인구유지도의의。
Three PCR methods(conventional PCR, touchdown PCR, nested PCR) were used to amplify the DNA topoisomeraseⅡfragments of Pleurotus ostreatus and P.eryngii var ferulae with degenerate prim-ers,which were designed with the gene sequences of DNA topoisomerase Ⅱfrom NCBI.Their PCR sensi-tivity and specificity were compared .The result showed that the sensitivity and specificity of nested PCR were highest , and its sensitivity was 100 fold higher than conventional PCR , which indicated that the nested PCR was more suitable for PCR amplification under degenerate primers .This study will have some guidance significance to obtain the specific gene through degenerate primers .
