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From 2012.igem.org

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-	Found a paper containing glucose and CO2 uptake rates.	+	Nisa
-	# Roelco J. Kleijn, Joerg M. Buescher, Ludovic Le Chat, Matthieu Jules, Stephane Aymerich, and Uwe Sauer, "Metabolic Fluxes during Strong Carbon Catabolite Repression by Malate in Bacillus subtilis," Journal of Biological Chemistry vol. 285(3), pp. 1587–1596, 2010	+
		+	Multihost vector (BBa_I742123) double digestion with EcoRI-SpeI and XbaI-PstI.
-	However, still missing the relation between the glucose uptake rate and temperature.	+	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).
-	Also missing rates for glutamine, potassium, oxygen.	+	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

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Revision as of 19:26, 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: 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