异硫氰酸胍法快速提取二球悬铃木组织总RNA的研究

针对二球悬铃木组织中多酚物质含量较高的特点,采用异硫氰酸胍法从二球悬铃木花序和叶片中提取到质量高、完整性好的总RNA,28S rRNA的亮度约为18S rRNA的2倍,通过RT-PCR克隆到二球悬铃木中与拟南芥Leafy基因同源的部分编码区。高质量的RNA为Northern杂交和利用同源序列法克隆二球悬铃木的相关基因提供了前提条件。

Guanidine Thiocyanate Method of Total RNA Isolation in Platanus aceriflia

Guanidine Thiocyanate method was used to isolate total RNA from the inflorescence tissue and leaves of Platanus aceriflia successfully with regard to high quantity of phenolic compounds in its tissue.The high-quality total RNA was obtained by guanidine thiocyanate that eliminated the interferences of polyphenols and acquired the 28S rRNA,which is approximately two times as light as the 18S rRNA.The isolated RNA was of sufficient quality for cloning partial cDNA of Leafy gene homologous to Leafy in Arabidopsis thaliana according to RT-PCR.The method described here was the basis of Northern blotting and cloning genes from P.aceriflia according to homologous sequence.