摘要
目的 当经典Trizol法提取骨组织RNA不能满足实验要求时,使用盐溶液再处理提取的一种改良方法.方法 对经典Trizol法提取的骨组织RNA,用异丙醇再沉淀时,加入柠檬酸钠、氯化钠混合盐溶液溶解,再抽提RNA.测定RNA纯度、浓度和完整性,聚合酶链反应（PCR）鉴定其有效性.结果 重新提取的RNA能够被焦磷酸二乙酯（DEPC）水充分溶解,盐溶液处理后的RNA浓度与处理前相近;处理后RNA的A260/A280比值、A260/A230比值分别为1.96 ±0.11、1.90 ±0.21,均分别显著高于处理前（1.52±0.05、0.27 ±0.08）;凝胶电泳示RNA完整性,逆转录后可扩增出目的基因.结论 改良Trizol法重提的总RNA纯度较常规法显著提高,且完整性好,可以满足进一步分子研究的要求.
Objective To introduce one method to improve the effect of bone RNA extraction when the classical Trizol method can not meet the experimental requirements.Methods The concentration,purity and integrity of the extracted RNA were tested.The RNA was reversely transcripted into cDNA,as a template which was then amplified to DNA.Results The re-extracted RNA could be fully dissolved by diethyl chlorophosphate （DEPC） water.This method could effectively reduce the interference of mucin carbohydrate.After the prevention of the sodium mixture solution,the concentration of the RNA was slightly lower than that before the prevention.The rate of A260/A280 and A260/A230 of RNA was both significantly higher afher the prevention than that before the prevention [（1.96 ± 0.11） vs.（1.52 ± 0.05）,（1.90 ±0.21） vs.（0.27 ±0.08） respectively].Agarose gel electrophoresis showed the RNA after prevention was complete and the polymerase chain reaction （PCR） procession could successfully amplify the targeted gene.Conclusion The improved extraction method could insure the purity,concentration and integrity of RNA,and meet the requirements for molecular research.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第9期1993-1995,共3页
Chinese Journal of Experimental Surgery
基金
江苏省研究生创新工程项目（CXZZ13_0834）
苏州大学青年预研基金项目（SDY2011A42）
