用低浓度硫氰酸胍提取高质量植物RNA

Preparation of High Quality Plant RNA With Low Concentration of Guanidinium Thiocyanate

目前分离ＲＮＡ的方法很多，但大多是基于动物材料建立的方法，而针对植物材料的方法并不多见．建立了一种从植物材料中分离高质量ＲＮＡ的硫氰酸胍／氯化锂／热酚法．与其他硫氰酸胍的方法相比，该方法所用硫氰酸胍的浓度仅为现有方法的１／４０（０１ｍｏｌ／Ｌ），降低了实验成本，且所得ＲＮＡ质量令人满意．用该方法分离的ＲＮＡ在琼脂糖凝胶电泳上可清晰分出４条核糖体ＲＮＡ（ｒＲＮＡ）带；用此ＲＮＡ进行ＲＮＡ印迹或分离ｍＲＮＡ进行体外翻译试验，均获得很好效果．

The guanidinium thiocyanateLiClhot phenol method for high purity and high integrity RNA isolation from plant tissues was established. Comparing with other methods that involved guanidinium thiocyanate, this method costs less and produces RNA molecules with better quality. The working concentration of guanidinium thiocyanate used was reduced more than 40 times compared with previous methods. The isolated RNA with this method gave 4 ribosomal RNA (rRNA) bands when analysed by formaldehyde agarose gel electrophoresis. Northern hybridization, mRNA isolation and following in vitro translation experiments performed with this RNA also gave good results.