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Team:Groningen/Notebook/Wetwork 6July2012
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Repeated cDNA synthesis of Bad_0307 O/N, kept OK samples in -80 freezer. | Repeated cDNA synthesis of Bad_0307 O/N, kept OK samples in -80 freezer. | ||
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Revision as of 17:02, 16 September 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).
Renske
Purified cDNA made on 05/07, using the same method as previously.
Concentrations on nanodrop (to be added):
- Fresh_0207_1: OK (~200)
- Fresh_0207_2: too low
- Bad_0207_1: too low
- Bad_0207_2:OK(~200)
- Bad_0307_1: too low
- Bad_0307_2: too low
Repeated cDNA synthesis of Bad_0307 O/N, kept OK samples in -80 freezer. Back to notebook
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