| 摘要 |
The present invention provides a method for amplifying mRNA in ultramicroquantity by approximately 10<8 >times which is possible to apply for generating a cDNA library, subtraction cloning, generating and analyzing a microarray, and analyzing a gene expression. After making mRNA of sample adsorbed to oligo (dT)-bound magnetic beads, a double-stranded cDNA is synthesized on magnetic beads, an antisense strand cDNA-bound magnetic beads are eliminated after adding a linker containing T7 promoter sequence at the 5' end, by using a sense strand cDNA in supernatant as a template and using an oligo (dT) primer wherein a linker containing SP6 promoter sequence is added, a double-stranded cDNA is synthesized again, the cDNA mixture is amplified by PCR using a known sequence at a linker part of the both ends of said double-stranded cDNA as a primer. In addition, sense strand/antisense strand cRNA is synthesized by T7 or SP6 polymerase using said cDNA mixture.
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