Team:Warsaw/Calendar-Main/20 May 2008

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(Difference between revisions)
Line 18: Line 18:
-
Annealing time: 4 minutes <br>
+
Elongation time: 4 minutes <br>
35 cycles <br>
35 cycles <br>
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Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion <br>
Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion <br>
-
Annealing temperature: 73&deg;C<br>
+
Elongation temperature: 73&deg;C<br>
Annealing time: 4 minutes <br>
Annealing time: 4 minutes <br>

Revision as of 15:12, 1 October 2008

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Michał K:
1.PCR - translation fusion: AID + T7 RNA-polymerase - optimalization (temperature gradient 60°C - 80°C).

Primers:

AIDlNrH and T7pXbSal
Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion
Elongation time: 4 minutes
35 cycles
2. Optimalization of PCR - translation fusion: AID + T7 RNA-polymerase - MgCl2 cocentration and number of cycles.

Primers:

AIDlNrH and T7pXbSal


Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion

Elongation temperature: 73°C
Annealing time: 4 minutes

3. Gel electrophoresis of PCR products.