莲雾果实高质量总RNA提取方法的建立

Comparison of Extraction Methods for High Quality RNA from Wax Apple(Syzygium samarangense Merr.et Perry)

为从莲雾果实中提取高质量RNA,以"黑珍珠"莲雾果实为材料,采用试剂盒法、十二烷基磺酸钠(sodium dodecyl sulfate,SDS)法、十六烷基三甲基溴化铵(hexadecyltrimethylammonium bromide,CTAB)法、改良CTAB法和Trizol法5种总RNA提取方法进行比较分析。结果表明,SDS法得到的总RNA质量较差,有严重的降解和DNA污染;Trizol法提取物中有较多蛋白和多糖等残留,无条带出现。CTAB法、改良CTAB法和试剂盒法均能获得质量较好的莲雾果实总RNA,经实时荧光定量聚合酶链反应验证得到条带与目的片段大小一致,表明该RNA均能满足后续分子生物学研究。

The objective of the study was to extract high quality RNA from wax apple（Syzygium samarangense Merr. et Perry） fruits. The effectiveness of five RNA extraction methods, i.e., Plant RNA Kit, hexadecyltrimethylammonium bromide（CTAB）, modified CTAB, sodium dodecyl sulfate（SDS） and Trizol, was evaluated. The results showed that the quality of total RNA extracted by SDS method was poor due to serious RNA degradation and genomic DNA contamination. The total RNA extracted with Trizol reagent contained various residues such as proteins and polysaccharides and exhibited no RNA bands. Plant Kit protocol, CTAB and modified CTAB methods were found to be suitable for total RNA extraction from wax apple fruit. RT-PCR analysis demonstrated that the RNA bands were consistent with the target fragment size, which showed that the extracted total RNA could be applied to subsequent molecular biology research.