Team:Groningen/Notebook/Wetwork 9July2012


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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

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

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