User:Karmella Haynes/Notebook/BioBrick cloning/2011/01/26

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01/26/11 ✓ Minipreps: SP1AB 3xmut (24) Minipreps > Check with E/P digests Reagent Volume Expected: 1-24. SP1AB 3x mut w.t. = 259, 317, 454, 587 site 1 = 454, 576, 587 site 2 = 317, 454, 846 site 3 = 259, 317, 1041 sites 1/2 = 454, 1163 sites 1/3 = 576, 1041 sites 2/3 = 317, 1300 sites 1/2/3 = 1617 15 μL/lane; 1% agarose DNA(plasmid) 2.0 μL 10X buffer 1.5 EcoRI 1.0 PstI 1.0 dH2O 9.5   15 μL --> 37°C/ ~15 min. --> Conclusions: Site #1 was not mutated at all. Keep clones 1-6 to confirm no off-target mutations, then use one good clone to mutate site#1. Order 2 more mut_SP1A primers