| 摘要 |
PROBLEM TO BE SOLVED: To provide a method for amplifying by 108 fold a very small amount of mRNA capable of being applied to the production of a cDNA library, a subtraction cloning, the production and analysis of a micro array and the analysis of a gene expression. SOLUTION: This method for amplifying the very small amount of mRNA comprises adsorbing the mRNA in a specimen on oligo (dt) bonded magnetic beads, synthesizing a double chain cDNA on the magnetic beads, adding a linker having a T7 promoter sequence at the 5' terminal of the cDNA, removing anti-sense chain cDNA-bonded magnetic beads, adding a linker having a SP6 promoter sequence by using a sense chain cDNA in the supernatant as a mold to form an oligo (dt) primer, synthesizing again the double chain cDNA by using the above primer, and performing a PCR by using a known sequence in the linker parts at the both ends of the double chain cDNA to amplify the cDNA mixture. Also, by using such cDNA mixture, the sense chain/anti-sense chain cDNA can be synthesized by using a T7 or SP6 polymerase. |
