User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/06/14

iGEM Project name 1

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Monday 14th June 2010 I performed 3 restrictions for the cI inverter as follows: Double restriction: Reactive............1x [ul] DNA..................5 BSA...................1 Buffer2.............2 H2O.................10 EcoR1...............1 PstI...................1 Total................20 Single restriction: Reactive..........1x [ul] DNA................5 BSA.................1 Buffer3...........2 H2O...............10 PstI.................2 Total..............20 Single restriction: Reactive...........1x [ul] DNA.................5 BSA..................1 Buffer4............2 H2O................10 EcoR1..............2 Total...............20 I also performed a PCR with rtth (hot start) to extract the cI inverter 1º Reaction Reactive...........................1x................4.x H2O...............................11.5ul...........46ul Buffer 3.3x......................5ul...............20ul Mg(OAc)2........................3ul...............12ul dNTP’s...........................2.5ul.............10ul Oligo up-suffix..............2.5ul.............10ul Oligo down-preffx.........2.5ul.............10ul DNA................................2ul.................8ul Total...............................30ul.............120ul 2º Reaction Reactive...............1x (ul)..........6x (ul) H2O.....................10.5............63 Buffer 3.3x............9................54 Rtth .....................0.5...............3 Total....................20...............78 I did 3 tubes for the cI inverter, negative and positive controls.