Dear DK, You are correct. we do not have any issue with IBs and its recovery. only when mixing the cell paste with lysis buffer to create a homogenous suspension for homogenisation we do not get uniform mass. moreover i wanted to know the reason for the slimy cell paste as we have taken about 7 trial batches with this particular clone. The first 4 batches were fine and we used to get compact cell mass. But recent batches show this slime or cell lysis. so was unable to understand the reason for this sudden change when all parameters of fermentation is unchanged.
But thanks for everybody's suggestion. On Mon, Nov 20, 2017 at 2:26 PM, DK <[email protected]> wrote: > In article <[email protected]>, Megha > Goyal <[email protected]> wrote: > >yes we also assume the same. so any suggestion how can we handle it. Our > >protein is in form of inclusion bodies and we get our protein but slimy > >cell mass makes it difficult to handle the cell paste. > > If it's in IBs then who and why cares about the "slime"? You'll need to > sonicate/French press anyway and that breaks up DNA nicely. I'd suggest > sonication (looooong sonication) after every pelleting/wash cycle. That's > the right thing to do when cleaning up IBs for refolding anyway. > > DK > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
