IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2014/07/30

iGEM Project name 1

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Week 7 Day 3 - Picked 1 white colony from 29/07/14 transformed dig-lig plates and added to 5 ml LB + 5 μl Ampicillin and incubated at 37 °C also added to 50 μl of PCR Master Mix containing: 1 μl DNTPs 2 μl Primer 1 2 μl Primer 2 5 μl Buffer 2 μl MgCl2 0.5 μl BIOTAQ Enzyme 37 μl Water - PCR programme: Step 1 : 95 °C 5 min Step 2 : 95 °C 1 min Step 3 : 54 °C 1.5 min Step 4 : 72 °C 1 min Step 5 : 72 °C 5 min Steps 2, 3, 4 repeated x 34 times - Run on a 1 % agarose TAE gel at 100V. Each well loaded with 2 μl of loading dye and 20 μl of DNA. Two 1kb ladders loaded. - DNA mini prep of cultures from 29/07/14 (DNA 2.0 level 0's and PRO, CDS, TER) (2.5 mls) - Glycerol stocks made from 1 ml of O/N culture. - Transformations of DNA iGEM #2-19 and Vectors #1-4 intro E.coli to generate more DNA. -Made 2L of LB agar. -Met with Laura Bowater to discuss the construction of the iGEM wiki pages. How we want it set out. What we have done already. What needs to be added. Ideas for layout, headings/categories, content. Discussed getting a dedicated camera/camera phone to take pictures within the lab. Set up an Instagram account and embed with the wiki pages. Pages to be put on the wiki: Human Practices Notebook Project Safety Attributions Judging Criteria Human Practices: GM - right or wrong? Policy makers - Letters to MP Food security Environment - Farmers, industries, bactericides Technology - Golden Gate Business Plan - Stakeholders: farmers, companies, LEDCs, scientists, Gov, interested public Contact InCrops and ADAPT (Low carbon group).