Imperial/Wet Lab/Lab Notebook/2007-09-18
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===Purification of LuxR-His=== | ===Purification of LuxR-His=== | ||
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*We have begun to clone out the mut3bGFP from a registry part into a his tag -vector to enable us to then purify the GFP later. Today we carried out PCR using olgios matching the restriction enzymes of the mut3bGFP in the registry. Using these oligos we successfully cloned out the mut3bGFP and we have begun to cut off the restruiction sites from the primers used. | *We have begun to clone out the mut3bGFP from a registry part into a his tag -vector to enable us to then purify the GFP later. Today we carried out PCR using olgios matching the restriction enzymes of the mut3bGFP in the registry. Using these oligos we successfully cloned out the mut3bGFP and we have begun to cut off the restruiction sites from the primers used. | ||
*Tomorrow we will prepare the vector and clone in the mut3bGFP | *Tomorrow we will prepare the vector and clone in the mut3bGFP | ||
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Latest revision as of 14:50, 9 October 2007
18 September 2007
Purification of LuxR-His
- A 100ml of medium has been incoculated with cells to be grown overnight. Tomorrow 20ml of these will be used to incolculate 1 litre of LB
- A small cultre of cells expressing LuxR have been lysed and the supernatent collected. A nickel column was run with this preliminary sample of the LuxR and these fractions are to be run out in a SDS-PAGE gel tomorrow
Purification of mut3bGFP
- We have begun to clone out the mut3bGFP from a registry part into a his tag -vector to enable us to then purify the GFP later. Today we carried out PCR using olgios matching the restriction enzymes of the mut3bGFP in the registry. Using these oligos we successfully cloned out the mut3bGFP and we have begun to cut off the restruiction sites from the primers used.
- Tomorrow we will prepare the vector and clone in the mut3bGFP
