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来源：百度知道 编辑：UC知道 时间：2024/05/09 18:53:09
辽宁新品系绒山羊毛囊兴盛期皮肤cDNA文库的构建及EST序列分析

摘要： SMART技术构建绒山羊（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， 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BlastX及B lastN软件进行同源对比，得到1200多条EST序列，本文从中选取20多条，利用tblast对其功能进行对比分析。
关键字：辽宁新品系 cDNA文库 EST序列

Construction of a Skin cDNA Library of Liaoning New-breeding Cashmere Goat and EST Sequence Analysis

Abstract: SMART technology to build 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, 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, PCR expansion with long-chain by full-length double-stranded cDNA. Diges

SMART technology to build 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, 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, PCR expansion with long-chain by 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 BlastX and B lastN software homologous comparison, received more than 1,200 of EST sequences, this paper selected from more than 20, using a comparative analysis of its function tblast .

Abstract: SMART technology to build cashmere go