通过基于特异性双链cDNA合成的消减去除已知的、丰富的cDNA物种。
Removal of known, abundant cDNA species by specific double-stranded cDNA synthesis-based subtraction.
作者信息
Yang X O, Huang X D, Zhao H L, Zhang J W
机构信息
National Laboratory of Medical Molecular Biology, Peking Union Medical College, Beijing, P.R. China.
出版信息
Mol Biotechnol. 1999 Jun;11(3):225-8. doi: 10.1007/BF02788681.
An efficient and simple method for removing known, abundant cDNA species from the cDNA pool of highly differentiated cells is reported. The method involves preparation of sscDNA, followed by dscDNA-synthesis of known, abundant cDNA species led by specific primers and removal of the synthesized dscDNA with hydroxyapatite. By using this method, the globin cDNAs were reduced to less than 10(-5) of their original abundance. The results suggest that this method may facilitate the isolation of new genes from specific cells or tissues.
报道了一种从高度分化细胞的cDNA文库中去除已知的、丰富的cDNA种类的高效简便方法。该方法包括单链cDNA的制备，然后以特异性引物引导已知的、丰富的cDNA种类进行双链cDNA合成，并用羟基磷灰石去除合成的双链cDNA。通过使用该方法，珠蛋白cDNA被减少到其原始丰度的不到10^(-5)。结果表明该方法可能有助于从特定细胞或组织中分离新基因。
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