[Establishment of studying system cloned vernalization-putative cDNA clone by means of differential display PCR].
Differential display PCR has been developed as a tool detecting and characterizing specially expressed gene in eukaryotic cells recently. In this paper, a studying system was established which applied the novel approach of differential display PCR to clone vernalization-related cDNA clone. Here methodological details were provided which included the purification of total RNA, removal of contaminated DNA, reverse transcription of cDNAs, parameters of PCR, electrophoresis of amplified cDNAs, recovery and reamplification of cDNAs. It was also identified a vernalization-putative cDNA clone (VPC) of VPC28 which was only expressed at the key stage of vernalization for 20 d in winter wheat. These results moreover made it readily applicable to a broad spectrum of similar studies.
Zhao
D Z
DZ
Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871.
Chen
M
M
Tan
K H
KH
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000302:isolation & purification; D014908:Triticum / Q000235:genetics*
D018076:DNA, Complementary; D012313:RNA