Team:Groningen/Notebook/Wetwork 6July2012

From 2012.igem.org

(Difference between revisions)
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Concentrations on nanodrop (to be added):
Concentrations on nanodrop (to be added):
-
Fresh_0207_1: OK (~200)
+
*Fresh_0207_1: OK (~200)
-
Fresh_0207_2: too low
+
*Fresh_0207_2: too low
-
Bad_0207_1: too low
+
*Bad_0207_1: too low
-
Bad_0207_2:OK(~200)
+
*Bad_0207_2:OK(~200)
-
Bad_0307_1: too low
+
*Bad_0307_1: too low
-
Bad_0307_2: too low
+
*Bad_0307_2: too low
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.

Revision as of 14:09, 14 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).


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.