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Team:Groningen/Notebook/Wetwork 7August2012
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Revision as of 16:54, 7 August 2012
Nisa
PCR of Fnr promoter and lacI promoter using Phusion.
PCR of amilCP (blue pigment) and amilGFP (yellow pigment) using Phusion.
PCR of lycopene and violacein using Phusion. Elongation time for this PCR took longer than the other PCR (~2 minutes) due to the size of the pigment coding region (~7 Kb)
Result:
- PCR of Fnr and LacI promoter worked, eventhough the fragments did not appear very bright and sharp on the agarose gel.
- PCR of amilCP and amilGFP worked perfectly. ~700 bp DNA fragments were shown on the agarose gel.
- PCR of lycopene and violacein showed unspesific binding (multiple DNA fragments shown on the agarose gel). There was probably a problem with the template that was used for lycopene PCR and higher annealing temperature is probably needed for violacein PCR.
However, PCR product(s) gained from these reactions (Fnr and LacI promoter, amilCP and amilGFP, lycopene from PCR on Saturday) were purified and used for further cloning steps.
