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 5t°C, 60°C and 65°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.