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Team:Freiburg/Notebook/TAL
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Revision as of 22:24, 25 September 2012
Our way to the TAL vector
08/08/12
1.) MutPCR for pSB1C3 vector backbone in order to remove restriction sites vor BsmBI
2.) Transformation of Sirt2 and HAT5
3.) GGC in order to produce our MammoBrick
1.) MutPCR:
| Tube 1 [µl] | Tube 2 [µl] | |
|---|---|---|
| Primer MutPCR fo [10 mM] | 1 | 0,25 |
| Primer MutPCR re [10 mM] | 1 | 0,25 |
| dNTPs [10 mM] | 1 | 1 |
| Phusion Buffer HF | 1,25 | 1,25 |
| Phusion Polymerase | 0,25 | 0,25 |
| DMSO | 1 | 1 |
| MgCl2 | 1 | 1 |
| Template | 1 | 1 |
| ddH2O | 5 | 6,5 |
Program for Thermocycler (20 cycles)
| 1. | 95°C | 5min |
| 2. | 95°C | 50s |
| 3. | 80°C | 50s |
| 4. | 72°C | 3min |
| 5. | 72°C | 7min |
| 6. | 4°C | // |
