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.


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