IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Gold Bioremediation/2010/07/15

iGEM Project name 1

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July 15th 2010 Digestion of pSB1A2 backbone, GolB, and GolS GolB+GolS 30 ul template 2 ul Buffer 1 ul XbaI 1 ul SpeI 16 ul dH2O pSB1A2 1.94 ul template 2 ul Buffer 1 ul XbaI 1 ul SpeI 44.06 ul dH2O note: Should have included BSA and added 5 ul Buffer for 50 ul reaction Gel Extraction of pSB1A2 No bands for GolS or GolB Gel extracted backbone using Zymo kit, got good concentrations. PCR-amplified GolB and GolS 41.1 ul H2O 0.5 ul template 1 ul Primer 1 1 ul Primer 2 1 ul dNTP 5 ul Buffer 0.4 ul polymerase 1. 95 degC for 5 min 2. 95 degC for 30 sec 3. 51 degC for 30 sec 4. 72 degC for 1 min 5. Repeat from step 2 for 31 cycles 6. 72 degC for 10 min