Team:Groningen/Notebook/Wetwork 6July2012

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Emeraldo/Tom
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1.) Plasmid isolation of pSac-CM. Conc: 40ng/ul
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2.) PalsT gradient PCR (50 degrees celcius until 60 degrees celcius) with different Primers: 150alsTF+Rev, 250alsTF+Rev, 300alsTF+Rev, and 500alsTF+Rev.
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3.) B. subtilis transformation using 2 methods (Molgen and team Edinburgh) with pSac-CM and BBa_I742123.
 
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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!!!
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5.) O/N E.coli with pSac-CM for another isolation (we used all of the isolated plasmid today. isolate more for further use).
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<p class="margin">Emeraldo/Tom<br>
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1.) Plasmid isolation of pSac-CM. Conc: 40ng/ul <br>
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2.) PalsT gradient PCR (50 degrees celcius until 60 degrees celcius) with different Primers: 150alsTF+Rev, 250alsTF+Rev, 300alsTF+Rev, and 500alsTF+Rev.<br>
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3.) B. subtilis transformation using 2 methods (Molgen and team Edinburgh) with pSac-CM and BBa_I742123.<br>
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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!!!<br>
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5.) O/N E.coli with pSac-CM for another isolation (we used all of the isolated plasmid today. isolate more for further use).<br>
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<br>
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<br>
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Renske<br>
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Purified cDNA made on 05/07, using the same method as [[Team:Groningen/Notebook/Wetwork_5July2012|previously]].<br>
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Concentrations on nanodrop (to be added):<br>
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*Fresh_0207_1: OK (~200)<br>
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*Fresh_0207_2: too low<br>
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*Bad_0207_1: too low<br>
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*Bad_0207_2:OK(~200)<br>
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*Bad_0307_1: too low<br>
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*Bad_0307_2: too low<br>
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Repeated cDNA synthesis of Bad_0307 O/N, kept OK samples in -80 freezer.<br><br>
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A></p><br><br>
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Latest revision as of 16:59, 25 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 [[Team:Groningen/Notebook/Wetwork_5July2012|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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