AFLP分子标记技术的改进.pdfVIP

生物技术通报 # # 2006 BIOTECHN OLOG Y B ULLETIN AFLP 刘平武 李赟 蔡强 宋传宇 何庆彪 杨光圣* (,, 430070) : 对AFLP技术的限制性内切酶组合进行了改进以两 个低频酶组合代替传统的高频酶与低频酶组 合进行 子标记筛选, 结果表明, 采用低频酶组合(E coR+P st)产生的条带比传统酶切组合 (E coR+M se和 Pst+M se)要少且 布不均匀, 多集中在 150~ 800bp之间, 条带信号强度要大, 多态性明显得到提高, 子标记 筛选成功率也得到了提高,而且其成本更低的特点有利于 AFLP技术在我国的进一步推广普及 : AFLP 子标记 限制性内切酶组合 改进 Improvem ent of Amplified Fragm ent L ength Polym orphism ( AFLP) Technique Liu Pingwu LiYun Cai iang Song Chuanyu He ingbiao Yang Guangsheng (NationalK ey Laboratory of Crop Genetic Imp rovem en,t H uazhon A ricultural Un ivers ity, R esearch Center of N ationalR ap eseed Imp rovem ent, Wuhan 430070) Abstract: In this study, AFLP ( amplified fragment length polymorphism) was mi proved on the combination of re- striction enzymes by using two average cutting frequency restriction enzymes (EcoR+P st) to take place the combina- tion of an average cutting frequency and a higher cutting frequency restriction enzyme (E coRI +M seI andP stI+M se I) in a single digestion. The results showed that the combination ofEcoR+P st could generate fewer fragmentswith larger size (most of bands distribute among 150 bp to 800bp) aswell as a higher percentage of polymorphism thanE coR+M se andP st+M se. The signal strength and polymorphism of PCR productswere mi proved too. Furthermore, the results described here have great commercial potential because the average cutt