Team:Groningen/Notebook/17 July 2009
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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. | 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! | :::→{{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=== | ===Transporters=== |
Revision as of 10:22, 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
Metal Accumulation
Vectors
Dry
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