Clifford W. Schweinfest , Kelly W. Henderson , Jian-Ren Gu , Stavros D. Kottaridis , Stavros Besbeas , Evi Panotopoulou , Takis S. Papas
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Subtraction hybridization cDNA libraries from colon carcinoma and hepatic cancer
cDNA clones of differentially expressed mRNAs in a colon carcinoma and a hepatocellular carcinoma have been isolated by subtractive cDNA cloning. The subtracted material is at least 90 × enriched for differentially expressed sequences and can be used for construction of subtractive cDNA libraries and polymerase chain reaction (PCR) amplification to generate differential probes. Commercially available λZAP II is used for construction of primary libraries since single-stranded phage bearing the cloned cDNA can be excised in vivo and because λ libraries are convenient for subsequent screening and manipulations. Rare mRNAs (less than 0.01% abundance), which are differentially expressed, can be isolated utilizing this procedure.
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