Team:Freiburg/Notebook/TAL

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Revision as of 16:36, 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] 10,25
Primer MutPCR re [10 mM] 10,25
dNTPs [10 mM] 11
Phusion Buffer HF 1,251,25
Phusion Polymerase 0,250,25
DMSO 11
MgCl2 11
Template 11
ddH2O 56,5


Program for Thermocycler (20 cycles)

1. 95°C5min
2. 95°C50s
3. 80°C50s
4. 72°C3min
5. 72°C7min
6. 4°C//