Team:Groningen/Notebook/17 July 2009

From 2009.igem.org

Revision as of 10:36, 17 July 2009

Wet

GVP Cluster

Discussion:

All colonies (transformants vector + promoter ligated with gvp) and o/n culture became red, so probably RFP is still in there... What might be the problem? The vector with promoter self-ligated due to uncomplete digestion (done 15 July 2009) leading to the following fragments:

• vector+RFP linear
• vector
• RFP

When the upper fragment (~2kb) is contaminated with linear vector this would lead high efficiency self-ligation instead of ligating gvp with the emtpy vector.

→TODO Therefore the restriction/purification/ligation will be redone and selection of non-red colonies will be made!

→DONE Also plasmid from the o/n cultures will be purified and analysed by restriction analysis.

• Used Sigma Plasmid isolation kit.
• DNA concentrations:

Transporters

PCR 16-07 Sample names ' ' ' ' ' ' ' ' ' Component B1 B2 C1 C2 A1 A2 E1 E2 F1 F2 MM 21 22 21 22 21 22 21 22 21 22 F 1uL Fw Fw Fw Fw mut1 mut1 Rev Rev Fw Fw R 1uL Rev Rev mut2 mut2 PCR2 PCR2 mut1 mut1 mut1RC mut1RC DNA 1uL 1 0 1 0 1 0 1 0 1 0 taq 1uL 1 1 1 1 1 1 1 1 1 1

Metal Accumulation

Vectors

Dry