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Team:Groningen/Notebook/Wetwork 9July2012
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| + | <p class="margin">Nisa<br> | ||
| + | Multihost vector (BBa_I742123) double digestion with EcoRI-SpeI and XbaI-PstI. <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. | + | 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> |
| + | <br> | ||
| + | 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> | ||
| - | 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. | + | 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> |
| + | <br> | ||
| + | Biobrick parts that was requested from iGEM HQ finally arrived!! <br> | ||
| + | <br> | ||
| + | List of biobrick parts:<br> | ||
| + | # BBa_K116603<br> | ||
| + | # BBa_K116602<br> | ||
| + | # BBa_J37019<br> | ||
| + | # BBa_K116639<br> | ||
| + | # BBa_K116609<br> | ||
| + | <br> | ||
| + | E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC<br> | ||
| + | <br> | ||
| + | <br> | ||
| + | Arjan<br> | ||
| + | Made cDNA of sample 0307_badmeat (duplo). O/N, 18 hours @42 degrees C.<br> | ||
| + | <br> | ||
| + | <A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A> | ||
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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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