trizol英文全称

由于SMART技术是利用逆转录酶内源的末端转移酶活性，只要单管，一步即可完成，不需要额外的cDNA抽提纯化或者沉淀，或者额外的酶反应，只需要少至25ng的mRNA或者50ng的Total RNA就可以得到高质量、高产量的cDNA库，更重要的是得到的cDNA能够代表原有样品中的mRNA的丰度，因此在本实验中把它直接用于构建绒山羊（Capra hircus）毛囊兴盛期皮肤组织的cDNA质粒文库。Trizol Reagent（GIBCO/BRL）分离RNA后，需要提取mRNA，这需要用Oligotex（QIAGEN），提取步骤如下：以锚定引物反转录合成cDNA第一链作为模板链，以两个锚定引物，用长链PCR扩增全长cDNA双链。用SfiⅠ酶切去除小片段后的CHROMA SPIN-400，连接到SfiⅠ消化的pBluescriptⅡ SK（带有SfiⅠA和B两个位点）质粒载体中，转化E.coli 5a，库容量为1.8×105clones。通过对文库中克隆子的序列测定和生物信息学初步分析，并且用B lastX及B lastN软件进行同源对比，得到1200多条EST序列，本篇文章就从中选取20多条进行分析，辽宁新品系绒山羊cDNA文库以及EST文库的成功构建,为研究辽宁新品系绒山羊功能基因和蛋白质组学奠定了基础。New Strain of Liaoning cashmere goat skin, hair follicles hugely popular for a cDNA library construction and EST sequence analysis Abstract: The SMART technology is the use of reverse transcriptase endogenous terminal transferase activity, as long as the single-tube, one step can be completed, no additional purification or precipitation of the cDNA extraction, or an additional enzymatic reactions, just as little as 25ng of the Total RNA or 50ng of mRNA can get high-quality, high-yield cDNA library, more importantly, to be able to represent the original sample of cDNA of mRNA abundance, so in this experiment used to construct it directly to cashmere goat (Capra hircus) follicle hugely popular skin tissue cDNA plasmid library. Trizol Reagent (GIBCO / BRL) separation of RNA, you need to extract the mRNA, which requires using Oligotex (QIAGEN), extraction steps are as follows: the anchor primer first-strand cDNA synthesis of reverse transcription as a template chain, with two anchor primers, using PCR amplification of long-chain full-length double-stranded cDNA. Digested with Sfi Ⅰ after the removal of small fragments CHROMA SPIN-400, connect to the Sfi Ⅰ digested pBluescript Ⅱ SK (with Sfi Ⅰ A and B 2 sites) plasmid vector and transformed into E.coli 5a, storage capacity of 1.8 × 105clones . Library clones by sequence determination and bio-informatics preliminary analysis, and with the B lastX and B lastN software homologous comparison, received more than 1,200 of EST sequences, this article will select from more than 20 analysis, Liaoning New strain cashmere goats EST library cDNA library as well as the successful construction of new lines for the study of Liaoning cashmere goats functional genomics and proteomics basis.