IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/09/21

iGEM Project name 1

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dspB Track Miniprep O/N cultures C,D,G,H Protocol: Rafael's Alkaline Lysis Protocol Resuspend DNA in 70uL of water Nanodrop: [C] = 1777.0ng/uL [D] = 1318.5ng/uL [G] = 1373.8ng/uL [H] = 842.5ng/uL Restriction Digest Protocol: "digest" protocol from biobrick Restriction Digest REAGENTS 1 RXN VOLUME (uL) Buffer 2 2 BSA 0.2 DNA 10 ddH20 7.8 Total 20 x = amount of DNA DNA: Cut terminator (suffix) at E/X Cut D and G at E/S Add 1uL of each appropriate enzyme Digest for 2 hours; start at 3:20pm Gel verification colony PCR products Protocol: gel verification protocol in Protocol (SOP) Machine conditions: 0.5x TBE buffer, 100V, 60min Gel orientation: Gel orientation D G 100bp ladder E/X term 1 kb ladder Results: Digestion failed. Re-Restriction Digest Protocol: "digest" protocol from biobrick Restriction Digest REAGENTS 1 RXN VOLUME (uL) Buffer 2 2 BSA 0.2 DNA 10 ddH20 7.8 Total 20 DNA: Cut terminator (suffix) at E/X Cut D and G at E/S Add 1uL of each appropriate enzyme O/N digestion starting at 9:30pm