title: Construction of a cDNA fragment library from SH-SY5Y cells using restriction display PCR.
abstract: A complementary DNA (cDNA) fragment library from SH-SY5Y cells is constructed using a restriction display polymerase chain reaction (RD-PCR) technique. Messenger RNA (mRNA) is extracted from SH-SY5Y cells and single-strand cDNA synthesised using an anchored oligo primer (dT18). The second strand is produced by nick translation. The double strands are cleaved with the restriction enzyme Sau3AI and the fragments ligated with universal linker. The products are amplified with universal primers and selected primers, ligated into the pMDI8-T vector, and then sequenced. The library constructed contained 136 subgroups, each comprising seven to 12 cDNA fragments. RD-PCR proved a simple, effective way to construct a cDNA library, and this will contribute to the investigation of gene expression in the neuron in future microarray studies.
authors: Institute of Molecular Biology First Military Medical University, Guangzhou, Peoples Republic of China.
authors: Zhang
authors: Z
authors: Bao
authors: Ma
authors: M
authors: Wenli
authors: Wu
authors: W
authors: Qinghua
authors: Guo
authors: G
authors: Qiuye
authors: Shong
authors: S
authors: Yanbin
authors: Zheng
authors: Z
authors: Wenling
mesh_terms: D018076:DNA, Complementary / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D004273:DNA, Neoplasm