[The construction of rapid amplification of cDNA ends cDNA libraries from human fetal bone and joint].
Xuan Liang, Yubao Gong, Qian-Ting Liu, Jianlong Li, B Chen, Chang-Jun Guo
Abstract
Xuan Liang, Yubao Gong, Qian-Ting Liu, Jianlong Li, B Chen, Chang-Jun Guo
Abstract
OBJECTIVE: To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone- and joint-specific development-related genes. METHODS: Total RNA of bone and joint were extracted with the modified single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. The double-stranded end-blunted cDNA were synthesized using TaKaRa's cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers. RESULTS: A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed. CONCLUSION: The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue-specific genes.
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OBJECTIVE: To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone- and joint-specific development-related genes. METHODS: Total RNA of bone and joint were extracted with the modified single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. The double-stranded end-blunted cDNA were synthesized using TaKaRa's cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers. RESULTS: A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed. CONCLUSION: The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue-specific genes.
Key concepts: Complementary DNA, Rapid amplification of cDNA ends, cDNA library, Biology, Molecular biology, Gene, Genetics, Molecular cloning