To determine specific mRNA without purification by adding a dissolution buffer and a hybridization buffer to a sample and reacting the sample with the labeled nucleic acid and fixing the sample to a vessel after reaction, adding a reverse transcriptase thereto and carrying out PCR reaction using a primer.
A dissolution buffer and/or hybridization buffer are added to a sample and as necessary, the sample is homogenized and the sample solution is brought into contact with labeled nucleic acid probe hybridized with RNA. A part of hybrid complex between the probe and mRNA is moved to a thermally stable reactor coated with an organic compound free from PAase and binding on the probe and the sample is incubated at about 4-50°C for ≥10sec. Then, the supernatant is removed from the reactor and the reactor is cleaned with a reaction buffer and a mixture containing a reverse transcriptase and a nucleotide is added and another mixture containing a thermally stable DNA polymerase and the specific primer and the nucleotide is added thereto and the mixture is amplified by PCR method to determine specific mRNA without purification.
HINTSUPEETAA MATEIASU
FURITSUTON HANSUUPEETAA
BUITSUTOORU HAIKO
