A MOTHED FOR QUANTIFYING GENE BASED POLYMERASE CHAIN REACTION

摘要

The invention involves molecular biological technology ,that is,a mothed for quantifying gene based polymerase chain reaction. In the invention, the mothed of PCR successive transcription reaction consists of two steps , the first step is producing PCR less 5-10 cycles than conventional PCR cycles, during which the amplification product enrichment in linear, and stopping before nonexponential amplification . Because a RNA polymerase promotor sequence is inserted to one of PCR primers at 5' terminal , PCR amplification product can be used as transcription template for RNA synthesis in vitro. The second step is promoting RNA synthesis as soon as adding transcription reagent, the yield of RNA synthesis is in proportion to PCR amplification product therefore can quantify primal template in the sample. The yield of RNA synthesis is multiplication hundred and thousand than template which can quantified simply using ethidium bromide or other nucleotide dye. The said gene quantitation method don ' t use radioactive isotope , don ' t involve antigen-antibody reaction , and don' t carrying out nucleotide hybridization . The said mothed is simple and accurate, and can be used broadly.