In vitro amplification of human basic fibroblast growth factor mRNA by RNA-specific exon-junction primers.
Highly sensitive and RNA-specific primers for the determination of human basic fibroblast growth factor (bFGF) gene expression by RT-PCR were identified. The RNA-specific primers could amplify bFGF mRNA from 10 pg to 1 ng of total cellular RNA without interfering with the presence of genomic DNA of the cell. The feasible temperatures of the primers annealed to the template were 55 degrees C, 60 degrees C and 65 degrees C. In addition, different locations of primers on the bFGF mRNA molecule yielded distinct amounts of RT-PCR products from the same concentration of RNA, suggesting that the mRNA secondary structure of bFGF affected the RT-PCR. Owing to high sensitivity and specificity of the primers to bFGF RNA, the RNA-specific primers may be potentially utilized for the determination of human bFGF gene expression by in situ RT-PCR.
Sheu
H M
HM
Department of Dermatology, National Cheng Kung University, Tainan, Taiwan.
Leung
M F
MF
Leung
W C
WC
Kuo
K W
KW
D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D012680:Sensitivity and Specificity
D018076:DNA, Complementary; C034477:RNA primers; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D012313:RNA