IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/16

UIUC - Arsenic Collection System!

Main project page Previous entry      Next entry

9/16/10 All overnights were white!!! 12:30 PM Miniprep Promega kit J23100B0034ArsRSDM2B0015pSB1T3 colony 1: 76.4 ng/μL colony 2: 75.9 ng/μL colony 3: 6.5 ng/μL colony 4: 50.2 ng/μL diluted colonies 1,2, and 4 to 20 ng/μL 1:30 PM Verification PCR Ingredient Amount Mastermix (5 reactions) 10X Buffer 2 μL 10 μL 10 mM dNTPs .4 μL 2 μL Template 1 μL (20 ng) added individually to each reaction F Primer .4 μL 2 μL R Primer .4 μL 2 μL Polymerase .2 μL 1 μL H2O 15.6 μL 78 μL Reaction for J23100B0034ArsRSDM2B0015pSB1T3 3 Ingredient Amount 10X Buffer 2 μL 10 mM dNTPs .4 μL Template 3.07 μL (20ng) F Primer .4 μL R Primer .4 μL Polymerase .2 μL H2O 13.53 μL Samples: J23100B0034ArsRSDM2B0015pSB1T3 1 J23100B0034ArsRSDM2B0015pSB1T3 2 J23100B0034ArsRSDM2B0015pSB1T3 3 J23100B0034ArsRSDM2B0015pSB1T3 4 negative control (H2O) Primers (10 micromolar): VF2 VR Program: 95°C 3 min 95°C 30 sec 55°C 30 sec 72°C 1 min go to step 2 30X 72°C 3 min hold at 4°C 5:20 PM Gel TAE, 1%, 175V Lane Sample Amount 1 1 Kb ladder 1 μL dye 2 J23100B0034ArsRSDM2B0015pSB1T3 1 1 μL dye, 5 μL sample 3 J23100B0034ArsRSDM2B0015pSB1T3 2 1 μL dye, 5 μL sample 4 J23100B0034ArsRSDM2B0015pSB1T3 3 1 μL dye, 5 μL sample 5 J23100B0034ArsRSDM2B0015pSB1T3 4 1 μL dye, 5 μL sample 6 negative control 1 μL dye, 5 μL sample Made glycerol stock of J23100B0034ArsRSDM2B0015pSB13 colony 1, ISB237