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Team:Groningen/Notebook/Wetwork 9July2012
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PCR of alsT promoter region using -150, -250, -300, -500 forward primer. | PCR of alsT promoter region using -150, -250, -300, -500 forward primer. | ||
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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. | 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. | ||
| Line 16: | Line 17: | ||
Biobrick parts that was requested from iGEM HQ finally arrived!! | Biobrick parts that was requested from iGEM HQ finally arrived!! | ||
| - | List of biobrick parts: | + | 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 | E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC | ||
Revision as of 19:28, 10 July 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
