Cell Lysis and Protein Extraction of MaSP2 9-mer and 12-mer constructs from BL21(DE3) cells
Followed using the protocol from Teule, et. al. 2012.
Lysis resulted in an extremely thick and viscous supernatant, with a clear yellow/tan color. Initial addition of deoxycholic acid to the cells seems very extreme; 1.5g/g of cells in lysis buffer. Results in a very thick paste forming; not quite sure if the membrane proteins are truly precipitating.
Additionally, a white, film/fibre like substance formed a layer on top of the lysate. Not quite sure exactly what this debris may be, but will run on an SDS PAGe gel tomorrow to analyze debris content.
Pellet saved after supernatant formation is viscous and snot like in nature; may possible be genomic DNA. Residual genomic DNA in the sample must be digested using DNase (3 uL total), to remove DNA from the solution before IMAC purification.