Method and Kit for enhanced differential display
    2.
    发明授权
    Method and Kit for enhanced differential display 失效
    用于增强差分显示的方法和工具包

    公开(公告)号:US5580726A

    公开(公告)日:1996-12-03

    申请号:US235180

    申请日:1994-04-29

    IPC分类号: C12Q1/68 C12P19/34

    摘要: An improved method for detecting and isolating differentially expressed mRNAs which comprises using first oligonucleotide primers for reverse transcription of mRNAs and both the first oligonucleotide primers and second oligonucleotide primers for amplification of the resultant cDNAs. The improvement of this method comprises providing first and second oligonucleotide primers with a length of at least 21 oligonucleotides. The method further comprises using a two-step PCR amplification, wherein non-stringent conditions are used for the first 1 to 4 cycles, and stringent conditions are used for the next 16 to 22 cycles. This highly reproducible method will permit the preparation of comprehensive catalogs of gene expression for any given cell type.

    摘要翻译: 一种用于检测和分离差异表达的mRNA的改进方法,其包括使用第一寡核苷酸引物用于mRNA的逆转录,以及用于扩增所得cDNA的第一寡核苷酸引物和第二寡核苷酸引物。 该方法的改进包括提供长度为至少21个寡核苷酸的第一和第二寡核苷酸引物。 该方法还包括使用两步PCR扩增,其中非严格条件用于前1至4个循环,并且在接下来的16至22个循环中使用严格条件。 这种高度可重复的方法将允许为任何给定的细胞类型制备基因表达的综合目录。