英语翻译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（Caprahircus）毛囊兴盛期皮肤组织的cDNA质粒文库.TrizolReagent（GIBCO/BRL）分离RNA后,需要提取mR

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英语翻译
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.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 .

Answer 1

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 .