"
Team:Groningen/Notebook/Modeling 9July2012
From 2012.igem.org
| Line 9: | Line 9: | ||
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. | ||
| | | | ||
| - | Result: | + | 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. |
| | | | ||
| | | | ||
Revision as of 13:30, 9 July 2012
(AP) 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: 1. BBa_K116603 2. BBa_K116602 3. BBa_J37019 4. BBa_K116639 5. BBa_K116609 | E. coli containing these parts were inoculated into LB agar+100ug/ml Ampicilin. Incubate overnight in 37oC.
