摘要
ＡＦＬＰ作为一种新的分子标记技术，其成功的关键依赖于ＤＮＡ的酶解效果，本文使用ＰｓｔＩ、ＴａｑＩ两种限制性内切酶，选用不同的酶切方式与不同的反应离子的组合对家蚕ＢＣ１代基因组ＤＮＡ进行酶切、通过人工接头连接、二次扩增及ＰＡＧＥ电泳、银染检测，结果显示是两酶在２５ｍＭＴｒｉｓ－Ａｃｅｔａｔｅ（ＰＨ７．８）、１００ｍＭＫＡｃ、１０ｍＭＭＧＡｃ２、１ｍＭＤＴＴ反应系统中能得到多态性丰富、重复性好的ＤＮＡ指纹图谱。同时，表明寻求适宜的限制酶缓冲组分是十分重要的。
AFLP (Amplified Fragment Length Polymorphism)is a new molecular marking technique. A successful AFLP is depended on the efficiency of endonuclease digesting DNA. Restriction endonucleases ,PstI and TaqI, was applied int applied int this experiment to excise the genomic DNA of Bombyx mori BC1 with various approaches of DNA digestion and ions.The result of silver staining indicated that PstI and TaqI was suitable in the reaction system at 25mMTris-Acetate (PH7.8)、 100mMKAc、10mMMgAc2、1mMDTT to get reliable genome polymorphism and powerful DNA fingerprinting map.It was also shown that composition of buffer must be researched in the reaction system.
出处
《江苏蚕业》
2000年第3期5-9,共5页
Jiangsu Sericul Ture
基金
江苏省科委
教委自然科学基金!(A1078和21075)
西南农业大学蚕丝学院开放课题基金资助
