[PCR walking in cotton genome using YADE method].
A Y-shaped adaptor dependent extension (YADE) method was developed to amplify the unknown genomic sequences adjacent to a known sequence, with the single-primer amplification completely suppressed. Using this method, two fragments, F027S and F027A, adjacent to a cotton ovule cDNA fragment (F027), were amplified using genomic DNA. It was demonstrated that F027S and F027A had 104 bp and 175 bp overlapped to F027, and contained 1 and 3 putative introns, respectively, all of which included conserved border sequence GT-AG and a putative branch site. The possible improvements and applications of YADE method are further discussed.
Xiao
Yue-Hua
YH
Biotechnology Research Center, Southwest Agricultural University, Chongqing 400716, China.
Luo
Ming
M
Fang
Wei-Guo
WG
Luo
Ke-Ming
KM
Hou
Lei
L
Luo
Xiao-Ying
XY
Pei
Yan
Y
D001483:Base Sequence; D018076:DNA, Complementary; D018744:DNA, Plant / Q000032:analysis; D018745:Genome, Plant; D003368:Gossypium / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods
D018076:DNA, Complementary; D018744:DNA, Plant