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人结肠平滑肌细胞提取物
来自 : mayitao
  • 货号:EH0066
    运输方式:干冰
    器官来源:消化系统
    欣润生物提供高质量的人、大鼠、小鼠即用型原代细胞提取物,包括基因组DNA、cDNA、总RNA、microRNA、总蛋白和细胞沉淀。我们提供的即用型原代细胞提取物可有效节约您为培养原代细胞并分离相应提取物所需的时间和精力。为确保可靠的实验结果,产品的数量和质量需经严格的质控测验。这些高质量的原代细胞提取物为细胞株试验的验证、各种生物医学数据的研究提供了强有力的工具。                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
    基因组DNA制备:采用酶解法裂解细胞,利用硅胶膜离心柱特异地吸附DNA,去除蛋白质、盐类、脂类等杂质,有效地保持基因组DNA的完整性。提取的DNA适用于酶切、PCR、SouthernBlot等实验。
    总RNA制备:利用QiagenRNeasyKit的Trizol试剂分离RNA,然后用QiagenRNACleanKit进行纯化。欣润生物提供的总RNA样品附有RNA样品OD值、浓度、总含量等。
    cDNA制备:纯化后的总RNA来合成cDNA第一链,以50ul总反应体系进行逆转录,体系中包括MMLV反转录酶和随机引物以及2μg总RNA。68℃反应10min后终止RT反应。利用1xRTBuffer运输cDNA,1μlcDNA足够做一次PCR反应。所有cDNA产品在订单发货之前都经过了严格的质控分析,保证了产品的质量。
    microRNA制备:采用具有超强的裂解能力和抽提灵敏度microRNA试剂盒制备,结合试剂盒采用的特殊吸附膜,大大增加了小分子RNA的吸附力,获得的microRNA纯度好,得率高,可以直接用于各种下游实验,如NorthernBlot分析,Real-TimePCR,MicroarrayAnalysis等。
    蛋白制备:利用改良型RIPABuffer(50mMTris,pH7.2,150mMNaCl,1mMEDTA,1mMEGTA,1%NP-40,1mMNa3VO4,5mMNaF,400uMPMSF,1ug/mlofPepstatinA,5ug/mlofAprotinin,5ug/mlofLeupeptin)制备细胞蛋白裂解液,离心去除细胞碎片,通过Bio-Rad蛋白检测试剂盒测定蛋白浓度,组织蛋白稀释后用非还原蛋白Buffer(50mMTris-HCl,pH6.8,5%Glycerol,1%SDS,0.002%BromphenolBlue)添加5%β-巯基乙醇及PMSF,-80度保存。
    细胞沉淀制备:首先分离培养出高质量的原代细胞,然后经平衡盐溶液清洗去除细胞表面杂质,最后用合适的裂解液裂解细胞并离心获得细胞沉淀。
    潜在的应用:<1>已知基因的PCR克隆;<2>mRNA选择性剪接;<3>基因克隆与目标测序;<4>WesternandNorthernBlot;<5>蛋白检测与蛋白质组学;<6>芯片研究与表达图谱分析。
     
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