IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/06/25

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6/25/10

12:40 PM

inoculated 5 mL broth with colonies from yesterday's biobrick plates. 37°C room to incubate

PCR

PCR of Plasmid Backbone
Component 20ul Rxn 6x Master Mix
10x Buffer 2ul 12ul
10mM dNTP --- ---
F Primer (52358135) 1ul (10uM) 6ul
R Primer (52358136) 1ul (10uM) 6ul
Template DNA (Plamsid Backbone) .5ul (~~250ng) (3)
Pfu Turbo DNA Pol 0.2ul 1.2ul
dH2O 15.3ul 91.8ul
PCR Program Used
95 degC 5 min
95 degC 30 sec 32 cycles
55 degC 30 sec 32 cycles
72 degC 270 sec 32 cycles
72 degC 5 min
4 degC hold


Miniprep of Parts for Backbones

Minipre of BB Plasmids for Backbone
Biobrick Part Backbone Concentration in ng/ul Label
RFP pSB1A2 430 A
RFP pSB1C3 513 C
RFP pSB1T3 428 T
E0034 pSB2K3 268 K

Gel

Lane Sample Amount
1. 1 Kb Ladder 3 ul
2. A 10 ul sample, 2 ul dye
3. C 10 ul sample, 2 ul dye
4. T 10 ul sample, 2 ul dye
5. K 10 ul sample, 2 ul dye
6. A' 10 ul sample, 2 ul dye
7. C' 10 ul sample, 2 ul dye
8. T' 10 ul sample, 2 ul dye
9. K' 10 ul sample, 2 ul dye
10. negative control 10 ul sample, 2 ul dye

Ran at 175V

10:10 PM

Moved inoculations from this afternoon (12:40) from 37°C room to 4°C room.


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