Wednesday, August 18, 2010

Insoluble protein?!

According to SDS P.A.G.E for solubility which I did on 18 AUG, the result doesn't looks good.

No PhaC protein on the soluble fraction; but a estimated ~15% expressed PhaC in insoluble fraction. [Troubleshooting: I still got other stock, will sonicate other bacterial cells and check again]

Maybe it is better to subclone the stored plasmid to Rosetta-gami2 (DE3) and BL21 (DE3) , BL21 (DE3)/pLySs?
At the same time, design primer and clone into pCold and pMAL.

Will email Dr after I have troubleshoot for insolubility and plan the cloning experiments in this weekend.

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