IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/09/02

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Abstract

  • Repeating the standardization

Main

Standardization

Digestion results

In the following electrophoresis gel (120V X 45min)

Lanes:

 1. Ladder 500 bp.
 2. PCR Digestion.
 3. pBBR1MCS-5 digestion.
 4. pBBR1MCS-5 plasmid
 The remaining lanes are not of interest for this analysis

Image:Sep_2_gel_shared_PGE.png

The PCR is not visible in this gel but a I think its not enough concetration to see it, however a problem could have arise during the PCR cleaning

Ligation

Regardless the problem of the PCR in the gel I did 3 ligations with these amounts:


Compound Rx 1 Rx 2 Rx 3 (control)
H20 3 3 6
Buffer T4 10x 1 1 1
Vector 1 2 2
PCR product 4 3 0
T4 DNA Ligase 1 1 1

I left the rections 5 hours

Transformation

I did the transformation of the previous ligations according to this protocol. At the end I had 4 petri dishes, three ligations and one medium control


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