IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/10/03

UIUC - Arsenic Collection System!

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10.3.10 10:30 diluted minipreps from yesterday to 20 ng/μL for PCR PCR Ingredient Amount Mastermix (10 reactions) 10X Buffer 2 μL 20 μL 10 mM dNTPs .4 μL 4 μL Template 20 ng (1 μL) added individually to each reaction F Primer .4 μL 4 μL R Primer .4 μL 4 μL Polymerase .2 μL 2 μL H2O 15.6 μL 156 μL Samples J2334A215C 1 J2334A215C 2 J2334A215C 3 J2334A215C 4 ArsRSDM2C 1 ArsRSDM2C 2 ArsRSDM2C 3 (+) control ArsR (-) control H2O Primers VF2 VR ArsRF EcoRI ArsR R Program 95°C 3 min 95°C 30 55°C 30 sec 72°C 90 sec go to step 2 30X 72°C 3 min hold at 4°C Gel: TAE 1% 175V Lane Sample Amount 1 1 Kb Ladder 1 μL 2 J2334A215 1 5 μL 1 μL dye 3 J2334A215 2 5 μL 1 μL dye 4 J2334A215 3 5 μL 1 μL dye 5 J2334A215 4 5 μL 1 μL dye 6 ArsRSDM2 1 5 μL 1 μL dye 7 ArsRSDM2 2 5 μL 1 μL dye 8 ArsRSDM2 3 5 μL 1 μL dye 9 (+) control ArsR 5 μL 1 μL dye 10 (-) control 5 μL 1 μL dye