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Dephosphorylation
Dephosphorylation
1. Precipitate DNA with 1/10 volume of NaOAC + 2 volume of EtOH or purified DNA through column.
2. Resuspend / elute DNA with 40 µl of DW then add to the reaction below.
DNA 40 µl
10x CIAP reaction buffer 5 µl
CIAP 0.6 µl
DW 5 µl
3. Incubate at 37 ◦C for 15 minutes then at 56 ◦C for another 15 minutes
4. Add another 0.6 µl of CIAP to the reaction
5. Repeat step 3.
6. Stop the reaction with 300 µl of stop-solution
10 mM Tris HCl (pH 7.5)
1mM EDTA (pH 7.5)
200 mM NaCl
0.5% SDS
7. Incubate at 75 ◦C for 10 minutes
8. Precipitate with 2 Volume of EtOH or purify through column
| Create Date : 28 มกราคม 2551 |
| Last Update : 28 มกราคม 2551 15:57:50 น. |
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If there is a stage at which an individual life becomes truly adult, it must be when one grasps the irony in its unfolding and accepts responsibility for a life lived in the midst of such paradox. One must live in the middle of contradiction, because if all contradiction were eliminated at once life would collapse. There are simply no answers to some of the great pressing questions. You continue to live them out, making your life a worthy expression of leaning into the light.
-Barry Lopez Arctic Dreams
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