TUDelft/29 September 2008

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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:</br></br> 1. 5' @ 94°C</br> 2. 1' @ 94°C</br> 3. 1' @ 65°C --> temperature is lowered with o.5°C per cycle</br> 4. 3' @ 72°C</br> 5. go to 2, 20 cycles in total</br> 6. 1' @ 94°C</br> 7. 1' @ 94°C</br> 8. 3' @ 72°C</br> 9. go to 6, 20 cycles in total</br> 10. 7' @ 72°C</br>