Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/12 August 2015"
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50C for 2 hr | 50C for 2 hr | ||
65C for 20 min | 65C for 20 min | ||
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Revision as of 06:45, 13 August 2015
08/12/2015
Colony PCR for M1 SwqAB 1 & 2 and M1-9(T7)
| 2X Taq MM | 6.25 uL |
| VF Primer | 0.625 uL |
| VR Primer | 0.625 uL |
| Template | 1 uL |
| ddH2O | 4 uL |
| 95C | 3 min |
| 95C | 30 sec |
| 72C | 25 sec (M1SeqAB)/ 1min 40 sec (M1-9) |
| x30 | |
| 72C | 5 min |
| 12C | Hold |
Grew liquid culture for
- all 4 colonies of M1-9 (T7); they all showed band close to the expected size ~1200-1300bp but they are slightly different so decided to sequence all of them.
- colonies 1-3 for M1SeqAB1
- none for M1SeqAB2 since we probably do not need this seq construct
BsaI Digestion of M1AB, BC and CA
| 1AB (553.88 ng/uL) | 1BC (1477.18 ng/uL) | 1CA (648 ng/ uL) | |
| Custmart | 5 uL | 5 uL | 5 uL |
| DNA | 7.2 uL | 2.7 uL | 6.2 uL |
| ddH2O | 33.8 uL | 38.3 uL | 34.8 uL |
| BsaI | 4 uL | 4 uL | 4 uL |
50C for 2 hr 65C for 20 min
