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Team:ETH Zurich/labblog/20140708
From 2014.igem.org
Contents |
Digest and miniprep
Tuesday, July 8th
Optimizing Restriction Endonuclease Reactions
Digest of 291 and 561
291: (2x)
1μL AscI
1μL FseI
0.5μL PvuI
0.5μL AseI
20 μL DNA 291
5μL Cut Smart Buffer
fill up with 22 μL H2O to 50μL
dephosphorylation of 181 and 271, cleanup with PCR clean-up system, gel electrophoresis
Gel electrophoresis
0.5 g Agarose in 50 mL TAE (0.5x), heat in microwave to dissolve
let solution cool down to approximately 50 °C, add 5 μL peqGREEN, mix
pour solution in tray, use appropriate comb
add 10μL loading dye (6x) to samples
fill samples and ladder (10 μL) in wells
run gel at 135 V for 50min
Miniprep of 35 and 36
