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TUDelft/29 September 2008
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(New page: =September 29th= ==Touchdown PCR== A touchdown PCR was performed with the new primers on the PCR products of atoB, idi and ispA and genomic DNA of E. coli. In both cases 60 ng of templat...) |
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==Touchdown PCR== | ==Touchdown PCR== | ||
| - | A touchdown PCR was performed with the new primers on the PCR products of atoB, idi and ispA and genomic DNA of E. coli. In both cases 60 ng of template DNA and the platinum Pfx enzyme was used. The PCR program looked like this:< | + | A touchdown PCR was performed with the new primers on the PCR products of atoB, idi and ispA and genomic DNA of E. coli. In both cases 60 ng of template DNA and the platinum Pfx enzyme was used. The PCR program looked like this:<br><br> |
| - | 1. 5' @ 94°C< | + | 1. 5' @ 94°C<br> |
| - | 2. 1' @ 94°C< | + | 2. 1' @ 94°C<br> |
| - | 3. 1' @ 65°C --> temperature is lowered with o.5°C per cycle< | + | 3. 1' @ 65°C --> temperature is lowered with o.5°C per cycle<br> |
| - | 4. 3' @ 72°C< | + | 4. 3' @ 72°C<br> |
| - | 5. go to 2, 20 cycles in total< | + | 5. go to 2, 20 cycles in total<br> |
| - | 6. 1' @ 94°C< | + | 6. 1' @ 94°C<br> |
| - | 7. 1' @ 94°C< | + | 7. 1' @ 94°C<br> |
| - | 8. 3' @ 72°C< | + | 8. 3' @ 72°C<br> |
| - | 9. go to 6, 20 cycles in total< | + | 9. go to 6, 20 cycles in total<br> |
| - | 10. 7' @ 72°C< | + | 10. 7' @ 72°C<br> |
Revision as of 08:46, 30 September 2008
September 29th
Touchdown PCR
A touchdown PCR was performed with the new primers on the PCR products of atoB, idi and ispA and genomic DNA of E. coli. In both cases 60 ng of template DNA and the platinum Pfx enzyme was used. The PCR program looked like this:
1. 5' @ 94°C
2. 1' @ 94°C
3. 1' @ 65°C --> temperature is lowered with o.5°C per cycle
4. 3' @ 72°C
5. go to 2, 20 cycles in total
6. 1' @ 94°C
7. 1' @ 94°C
8. 3' @ 72°C
9. go to 6, 20 cycles in total
10. 7' @ 72°C
