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采用异硫氰酸胍/氯化铯超离心法,从人脑恶性胶质瘤手术标本中抽提总 RNA;经两轮Oligo(dT)—cellulose 亲和层析纯化得到 mRNAs。Northern 转移印迹分析表明:mRNAs 无降解,完整性好。以此为模板,经反转录,合成得到双链 cDNAs 分子。cDNA 第1条链的反转录效率为36.4%,第2条链的合成效率为80.8%。
Total RNA was extracted from human malignant glioma surgical specimens using guanidinium isothiocyanate / cesium chloride ultracentrifugation; mRNAs were purified by two rounds of Oligo (dT) -cellulose affinity chromatography. Northern blot analysis showed that: mRNAs without degradation, integrity is good. Using this as a template, double-stranded cDNAs were synthesized by reverse transcription. The reverse transcription efficiency of the first strand of the cDNA was 36.4%, and that of the second strand was 80.8%.