The fiber length trait (FLT) of 538 individuals from nature birch population in Maorshan region, Heilongjang, China were measured, of which 100 individuals were selected as representative variety of correlated fragments screening with random amplified polymorphism DNA (RAPD) technique. In total of 20 RAPD primers were tested through multiple regression analysis between amplified strip and the character behaviors, and a correlative segment BFLR-16 was obtained. The correlation coefficient between BFLI-16 and FLT was 0.6144, with the significant level of 1%. BFLI-16 was then cloned, sequenced and transformed into SCAR marker. The percentage of identifying long fiber birches by this SCAR was more than 92. The result indicates that the SCAR markers has high specificity for the long fiber individuals and is highly linked with the gene controlling the character of fiber length, and its existence is significantly correlative with the increase in the fiber length.
以不同白桦个体为材料,通过琼脂糖凝胶电泳法和聚丙烯酰胺凝胶电泳法分析了DNA提取、双酶切、连接、预扩增和选择性扩增过程中的各影响因素,建立了扩增酶切片段长度多态性(Amplified Fragment Length Polymorphism)分析体系,并用E2+M4引物对不同白桦个体进行了扩增,得出优化的关键因素分别为:模板DNA的质量浓度为0.094g/L和0.086g/L,符合AFLP分析中的双酶切要求;酶切反应时间为2h15min;连接最适反应时间为2h;预扩增PCR的退火温度设为60℃。银染结果表明:扩增信号强,无背景干扰,条带清晰。