User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/06/17

{| width="800"
 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] iGEM Project name 1
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Thursday 17th June 2010
I performed a hot start PCR to the Green Fluorescent Protein with LVA tag for rapid degradation (BBa_K145015) to add an RBS (BBa_B0034) and 7 bases before the initial ATG, for this I used rtth polymerase.


 * 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).........4x (ul)

H2O.....................10.5............42

Buffer 3.3x............9................36

Rtth.....................0.5...............2

Total....................20...............80

The PCR came out well.


 * }