IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/08/27

From OpenWetWare

Jump to: navigation, search
iGEM Project name 1 Main project page
Previous entry      Next entry

Abstract

  • Repeating the standardization


Main

Standardization

PCR result

The result of the yesterday's PCR are shown in the following electrophoresis gel (120V X 35 min):

Lanes:

 1. Ladder 500bp.
 2. PCR

The band corresponds to the correct size (~1.5 kb) and is clean enough to further experiments.

PCR and plasmid digestion

Before I did the PCR product digestion, I cleaned up with the purification kit by Roche

In order to do the ligation by sticky ends I also did the double digestion by apaI and sacI of both, the pBBRMCS-5 plasmid and the PCR of the RFP.

The reactions were done with following compounds:

RFP PCR digestion

Compound μL
H20 3
DNA 10
Buffer 4 2
BSA 2
sacI 1.5
apaI 1.5
Total 20

pBBR1MCS-5 digestion

Compound μL
H20 5
DNA 7
Buffer 4 2
BSA 2
sacI 1.5
apaI 1.5
Total 20



Personal tools