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

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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.