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目的介绍一种快速简便地同时建立基因组DNA库和cDNA库的方法及详细步骤,并强调计算机管理的重要性。方法采5ml外周血,其中4ml分成2份用0.2%氯化钠处理收集白细胞,再用基因组DNA提取试剂盒抽提基因组DNA;另1ml血分成4份用直接裂解全血的方法提取RNA和一步合成法进行cDNA的合成。结果用该方法建立的库中每个基因组DNA样本平均可以做1200次PCR反应,每个cDNA样本平均可以做190次荧光定量PCR反应。结论对同一患者,用该方法可以既保存基因组DNA又保存cDNA,而且方法简便、实用,加上合理的计算机管理,可以快速准确地从众多的DNA中查找到自己需要的样品。
OBJECTIVE: To introduce a rapid and simple method and detailed procedure for simultaneous establishment of genomic and cDNA libraries and emphasize the importance of computer management. Methods 5ml peripheral blood was collected, of which 4ml was divided into 2 parts treated with 0.2% sodium chloride to collect leucocytes, and then genomic DNA extraction kit for genomic DNA extraction. Another 1ml of blood was divided into 4 parts by directly lysing whole blood RNA and a step Synthesis of cDNA synthesis. Results On average, 1,200 PCR reactions were performed on each genomic DNA sample set up by this method. The average fluorescence intensity of each cDNA sample can be measured 190 times. Conclusion This method can save both genomic DNA and cDNA in the same patient, and the method is simple and practical. With reasonable computer management, you can quickly and accurately find the samples you need from numerous DNA samples.