条斑紫菜丝状体总RNA提取方法比较

Comparison of Extraction Methods of Total RNA from Conchocelis in Porphyra yezoensis

目的:为了获得质量较高的条斑紫菜丝状体总RNA,对几种常用提取方法进行研究。方法:以条斑紫菜自由丝状体为材料,比较了用异硫氰酸胍法、CTAB法、SDS/酚法、TRIzol法、RNAplant法提取的RNA的质量和纯度。结果:异硫氰酸胍法提取RNA的成本低,但纯度不高;CTAB法产率较小,且不能完全去除多糖或蛋白质;SDS/酚法未能获得完整的RNA;TRIzol法未能见到5SrRNA条带,且带有杂带;而RNAplant法提取RNA的质量好、纯度高、提取效率高,其D260nm/D280nm值为1.836,经逆转录得到的双链cDNA扩增产物长度在200bp以上。结论:实验结果表明RNAplant法更适于条斑紫菜丝状体总RNA的提取。

Objective： In order to develop a simple and efficient protocol for isolating total RNA from conchocelis of Porphyra yezoensis, several methods were investigated. Methods： Guanidine thiocyante method, CTAB method, SDS/hydroxybenzene method, TRIzol method and RNAplant method were employed to isolate and purify the total RNA from the free living conchocelis of P.yezoensis. The quality and quantity of RNA were compared among these methods. Results： The RNA extracted by guanidine thiocyante method had low expense but poor purity; the RNA extracted by CTAB method had poor yield and remained some protein and polysaccharide; the RNA extracted by SDS/hydroxybenzene method lost some of RNA; the RNA extracted by TRIzol method didn＇t have high quantity; while the RNAplant method produced RNA with good quality and high purity. The value of D260nm/D280nm was 1,836. The double strand cDNAs synthesized by reverse transcription were larger than 200 bp. Conclusion： The results show that RNAplant method is fit for the isolation of the total RNA from the conchocelis of P.yezoensis.