Title: Construction of renal cell carcinoma cDNA subtractive library and cloning of renal cell carcinoma relative genes
Abstract: Objects [WT5BZo constuct a cDNA sustractive li brary of human renal cell carcinoma (RCC) with technique called suppression subt ractive hybridization. [WT5HZ]Methods Poly (A)+ RNA were isolate d form tissues of RCC and normal kidney respectively.Moreover, double-strand cD NAs were synthesized in turn. RCC cDNAs then were restricted and divided into tw o groups and ligated to the specific adaptorl and adaptor2 respectively .After R CC cDNAs hybridized with normal kidney cDNA twice and underwent two times of nes ted PCR ,then with arms of T/A plasmid vectors to set up the subtractive library . Results Human RCC subtrctive library with high subtract ive efficiency was set up sucessfully. The amplified library contains 350 positi ve clones. 95%of clones contain 50-400bp inserts.Conclusions The cDNA subtracti ve library of human RCC is a highly efficient one and lays solid foundation for screening and cloning new and specific oncogenes or tumor suppressor genes of RC C.
Publication Year: 2001
Publication Date: 2001-01-01
Language: en
Type: article
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