PMID- 8687428 OWN - NLM STAT- MEDLINE DCOM- 19960816 LR - 20061115 IS - 0006-291X (Print) IS - 0006-291X (Linking) VI - 223 IP - 3 DP - 1996 Jun 25 TI - Relative ratios of mRNA molecules encoded by genes with homologous sequences using fluorescence-based single-strand conformation polymorphism analysis. PG - 520-5 AB - The combination of reverse transcribed (RT)-PCR and fluorescence-based single-strand conformation polymorphism (SSCP) analysis is proposed for the quantitative determination of the ratio of mRNA molecules with homologous sequences. We applied this procedure to lactate dehydrogenase (LDH) subunits A and B. We designed fluorescence labeled common PCR primers in the sequences highly homologous between LDH-A and LDH-B cDNAs and performed RT-PCR-SSCP analysis. When PCR efficiency was almost the same between the different target sequences, analysis of mixtures of known amounts of LDH-A and LDH-B revealed linear and precise proportions of LDH-A mRNA. Template concentrations and PCR cycles did not affect the determination of proportions of LDH-A to total LDH. The present procedure could be easily applied to investigation of expression levels of genes encoding mRNAs with homologous sequences. FAU - Maekawa, M AU - Maekawa M AD - Division of Clinical Laboratory, National Cancer Center Research Institute, Tokyo, Japan. FAU - Sugano, K AU - Sugano K FAU - Ushiama, M AU - Ushiama M FAU - Masuda, T AU - Masuda T FAU - Ohkura, H AU - Ohkura H FAU - Kakizoe, T AU - Kakizoe T FAU - Sekiya, T AU - Sekiya T LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Biochem Biophys Res Commun JT - Biochemical and biophysical research communications JID - 0372516 RN - 0 (DNA Primers) RN - 0 (Macromolecular Substances) RN - 0 (RNA, Messenger) RN - EC 1.1.1.27 (L-Lactate Dehydrogenase) SB - IM MH - Amino Acid Sequence MH - Base Sequence MH - Cell Line MH - Colorectal Neoplasms MH - DNA Primers MH - Exons MH - Fibroblasts MH - Humans MH - L-Lactate Dehydrogenase/*biosynthesis/*genetics MH - Macromolecular Substances MH - Molecular Sequence Data MH - Pancreatic Neoplasms MH - Polymerase Chain Reaction MH - *Polymorphism, Single-Stranded Conformational MH - RNA, Messenger/biosynthesis/*metabolism MH - Stomach Neoplasms MH - Templates, Genetic MH - Tumor Cells, Cultured EDAT- 1996/06/25 00:00 MHDA- 1996/06/25 00:01 CRDT- 1996/06/25 00:00 PHST- 1996/06/25 00:00 [pubmed] PHST- 1996/06/25 00:01 [medline] PHST- 1996/06/25 00:00 [entrez] AID - S0006-291X(96)90927-4 [pii] AID - 10.1006/bbrc.1996.0927 [doi] PST - ppublish SO - Biochem Biophys Res Commun. 1996 Jun 25;223(3):520-5. doi: 10.1006/bbrc.1996.0927.