Matt Gethers/CRI, Thailand/Labwork/Digests/PstI
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PstI
Reaction Mix
| Reagent | Volume |
| Template | 10 μl |
| PstI | 0.5 μl |
| Fast Digest Buffer | 2 μl |
| H20 | 7.5 μl |
| Total | 20 μl |
Reaction Conditions
Incubate mixture at 37oC for 30 minutes and inactivate at 65oC for 15 minutes on the heat block.
Run Notes
8.12.08
Digesting pIs001.P1, P9a, P9b, P10a, P10b with PstI to determine if HmgR ORF is present. P1 is my positive control. I made a master mix with 63.75 μl water, 10.2 μl buffer, and 2.55 μl PstI. Aliquotted 15 μl of this master to each of 5 tubes, then added 5 μl of each template. Incubated at 37 degrees for 15 minutes and then loaded immediately on gel (no inactivation). Gel results here.
