Team:Groningen/Notebook/Wetwork 9July2012

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Nisa
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Multihost vector (BBa_I742123) double digestion with EcoRI-SpeI and XbaI-PstI.
 
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Result: Gel image of this vector is inconsistent. BBa_I742123 was isolated from 3 different E. coli colonies and was double-digested. The result of this reaction, however, varies between isolates (gel picture will be updated soon).
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PCR of alsT promoter region using -150, -250, -300, -500 forward primer.
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<p class="margin">Nisa<br>
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Last week the annealing temperature of each primer sets has been optimized. We experienced the best result in 58-60oC. Today the PCR was done with different polymerase, Pfu, instead of Taq. Annealing temperature used in this PCR was 60oC.
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Multihost vector (BBa_I742123) double digestion with EcoRI-SpeI and XbaI-PstI. <br>
-
Result: Only primer pair (Fwd-150,Rev) resulted in DNA fragment in the 0.8% agarose gel. The other primer pairs failed to give the expected results.
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Result: Gel image of this vector is inconsistent. BBa_I742123 was isolated from 3 different E. coli colonies and was double-digested. The result of this reaction, however, varies between isolates (gel picture will be updated soon).<br>
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<br>
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PCR of alsT promoter region using -150, -250, -300, -500 forward primer.<br>
 +
<br>
 +
Last week the annealing temperature of each primer sets has been optimized. We experienced the best result in 58-60oC. Today the PCR was done with different polymerase, Pfu, instead of Taq. Annealing temperature used in this PCR was 60oC.<br>
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Biobrick parts that was requested from iGEM HQ finally arrived!!  
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Result: Only primer pair (Fwd-150,Rev) resulted in DNA fragment in the 0.8% agarose gel. The other primer pairs failed to give the expected results.<br>
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<br>
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Biobrick parts that was requested from iGEM HQ finally arrived!! <br>
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<br>
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List of biobrick parts:<br>
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# BBa_K116603<br>
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# BBa_K116602<br>
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# BBa_J37019<br>
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# BBa_K116639<br>
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# BBa_K116609<br>
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<br>
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E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC<br>
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<br>
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<br>
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Arjan<br>
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Made cDNA of sample 0307_badmeat (duplo). O/N, 18 hours @42 degrees C.<br>
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<br>
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
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List of biobrick parts: 1. BBa_K116603 2. BBa_K116602 3. BBa_J37019 4. BBa_K116639 5. BBa_K116609
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{{Template:SponsorsGroningen2012}}
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E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC
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Latest revision as of 17:03, 25 September 2012







Nisa
Multihost vector (BBa_I742123) double digestion with EcoRI-SpeI and XbaI-PstI.
Result: Gel image of this vector is inconsistent. BBa_I742123 was isolated from 3 different E. coli colonies and was double-digested. The result of this reaction, however, varies between isolates (gel picture will be updated soon).

PCR of alsT promoter region using -150, -250, -300, -500 forward primer.

Last week the annealing temperature of each primer sets has been optimized. We experienced the best result in 58-60oC. Today the PCR was done with different polymerase, Pfu, instead of Taq. Annealing temperature used in this PCR was 60oC.
Result: Only primer pair (Fwd-150,Rev) resulted in DNA fragment in the 0.8% agarose gel. The other primer pairs failed to give the expected results.

Biobrick parts that was requested from iGEM HQ finally arrived!!

List of biobrick parts:
# BBa_K116603
# BBa_K116602
# BBa_J37019
# BBa_K116639
# BBa_K116609

E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC


Arjan
Made cDNA of sample 0307_badmeat (duplo). O/N, 18 hours @42 degrees C.

Back to notebook



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