改良TRIzol法高效提取钉螺组织总RNA

Extracting total RNA from Oncomelania snails by improved TRIzol technique

目的解决钉螺组织中总RNA提取难度大的问题,建立一套适合钉螺组织总RNA提取方法。方法采用改良TRIzol法从钉螺组织中提取其总RNA,并与传统TRIzol法提取结果进行比较。结果本方法优于传统TRIzol法,能够在钉螺组织中提取出完整性好、纯度高的总RNA,其A_(260)/A_(280)在1. 9～2. 0之间,A_(260)/A_(230)在1. 8～1. 9之间,电泳结果表明获得总RNA 28S、18S及5S条带清晰完整,无弥散现象;同时,选取β-acting基因检测,电泳结果表明所得总RNA的质量符合钉螺RT-PCR检测的实验要求。结论本方法便于实施,重复性好,适合钉螺组织总RNA提取,同时为改良优化传统TRIzol法提取其他医学贝类组织总RNA提供了研究思路。

Objective To develop an efficient technique for extracting total RNA from Oncomelania snails.Methods Improved TRIzol reagent was used to extract total RNA from Oncomelania hupensis,and the output was compared with conventional TRIzol procedure.Results Improved TRIzol reagent resulted in full and purified total RNA of snails,with A260/A280 and the A260/A230 ratio ranging from 1.9~2.0 and 1.8~1.9,respectively.Sub-unit RNA 28S,18S and 5S were clearly and completely generated on the electrophoretogram without any dispersion.Further verification indicated that the extracted total RNA was in high quality capable of detectingβ-acting gene by RT-PCR.Conclusion Improved TRIzol technique can be easy in performance and reproducible for extraction of total RNA from Oncomelania snails.Also,this technique may be a new concept for extracting total RNA from similar shellfish associated with medical importance through conventional TRIzol reagent.