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Revision as of 19:01, 10 July 2012

Emeraldo/Tom

1.) Plasmid isolation of pSac-CM. Conc: 40ng/ul

2.) PalsT gradient PCR (50 degrees celcius until 60 degrees celcius) with different Primers: 150alsTF+Rev, 250alsTF+Rev, 300alsTF+Rev, and 500alsTF+Rev.

3.) B. subtilis transformation using 2 methods (Molgen and team Edinburgh) with pSac-CM and BBa_I742123.

4.) pSac-CM restriction with different enzymes to confirm the stability of the plasmid. Enzymes used: ScaI-EcoRV-HindIII, NcoI-EcoRI-SmaI, and single digest using EcoRI-XbaI-SpeI-PstI. Result: the plasmid is OK!!!

5.) O/N E.coli with pSac-CM for another isolation (we used all of the isolated plasmid today. isolate more for further use).