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Fwd: Slimy recombinant e.coli pellet

Megha Goyal via methods%40net.bio.net (by mgbiotec from gmail.com)
Mon Nov 20 22:46:31 EST 2017


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 <dk from no.email.thankstospam.net> wrote:

> In article <mailman.123.1510862205.15473.methods from net.bio.net>, Megha
> Goyal <mgbiotec from gmail.com> 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
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