Title: Construction and identification of a differentially expressed genes subtracted cDAN library of human dermal papillae cells with aggregative behavior
Abstract: Objective To construct a differentially expressed genes subtracted cDNA library of dermal papillae cells(DPC) with aggregative behavior Methods Total RNA was extracted from DPC with and without aggregative behavior and double strand cDNA was synthesized by using SMART cDNA synthesis, respectively The cDNA fragments of differentially expressed genes in dermal papillae cells with aggregative behavior were isolated by suppression subtractive hybridization and then directly inserted into T/A cloning vector to set up the subtracted library Verification of the insert fragment was performed on 100 positive clones randomly screened by PCR method and among which 30 clones with insert fragments were verified by cDNA dot blot Results The amplified library contained approximately 320 positive bacteria clones Random analysis of 100 clones with nested PCR method showed that 95% clones contained 100~600 bp inserts cDNA dot blot showed that 27 clones (90%) were positive Conclusion A differentially expressed genes subtracted cDNA library of dermal papillae cells(DPC) with aggregative behavior is constructed by suppression subtractive hybridization combining with SMART cDNA synthesis The library is a highly efficient one and lays solid foundation for screening and cloning new and specific genes related to aggregative behavior of DPC
Publication Year: 2003
Publication Date: 2003-01-01
Language: en
Type: article
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