利用Y-RACE法进行棉花胚珠cDNA末端快速扩增

在YADE(Y shapedadaptordependentextension)方法的基础上 ,设计了一种新的cDNA末端快速扩增方法 ,称为Y RACE法 .利用该方法只需一次cDNA合成就能进行多个基因的 3′和 5′末端的扩增 .分别利用Y RACE方法和RACE试剂盒 (TaKaRa)扩增了一个棉花胚珠cDNA片段F0 2 7的末端序列 .序列比较表明 ,用Y RACE方法扩增的末端序列较试剂盒所得序列在最末端稍短 ,但同样能进行正确拼接并获得全长编码序列 .对Y RACE法的优缺点和可能的改进作了进一步讨论 .

Rapid Amplification of Cotton Ovule cDNA Ends by Y-RACE

Based on the Y shaped adaptor dependent extension (YADE) method, a new RACE method, designated as Y RACE, was developed. With this method, the 3′and 5′terminals of most of cDNA species present in the sample can be amplified from a single reaction of cDNA synthesis. The 3′and 5′cDNA ends of a differential cDNA AFLP fragment F027 from cotton ovules were amplified by Y RACE and RACE kits (TaKaRa). Sequence alignment demonstrated that the two fragments obtained by Y RACE, although several base shorter in the ultimate terminal than those amplified by RACE kits, could be joined to gain the complete coding region. The possible improvements and applications of Y RACE method were further discussed.