er to:
Tan, S. , et. al JMB 2000,297(4), 947-59.
Thanks,
Debanu.
From: CCP4 bulletin board on behalf of bputcha
Sent: Mon 7/16/2007 9:01 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Protein-DNA complex for crystallization
Hi,
I am trying to crystallize a protei
Hi Kumar,
1) While you contemplate other ideas, have you tried the following already?
- microseeding or macroseeding with the tiny crystals you have?
- crystallization using sitting drop vapour diffusion under oil? Often, one
gets a burst of
nucleation with several tiny crystals. Not sure if thi
On Jul 16, 2007, at 12:01 PM, bputcha wrote:
I am trying to crystallize a protein-DNA complex. I purify the
protein finally
using gel filtration. I purchase
single stranded complementary oligos (desalting from idtdna.com),
mix them up
How can I purify the duplex DNA further?
in my exper
ssouri.edu/TannerGroup/tanner.html
> From: <[EMAIL PROTECTED]>
> Reply-To: <[EMAIL PROTECTED]>
> Date: Mon, 16 Jul 2007 11:26:42 -0500
> To:
> Subject: Re: [ccp4bb] Protein-DNA complex for crystallization
>
> Dear Kumar,
> One often has to try duplexes with many differ
Kumar and Joe-
I'm not sure if this helps but I always purified my oligos prior to
crystallization. Originally, I would order them with the trityl-group on
and purify then on a reverse phase HPLC column and used TFA to cleave on
column. Although towards the end of the project, I just ordered the
Dear Kumar,
One often has to try duplexes with many different
ends before getting decent crystals (e.g. 18 for
one project in my lab, even more for others).
Depending on your Kd, you might find that your
complex falls apart during gel filtration.
How long are your oligos? Gel purification
som
Hi Kumar,
I also have the same issue. If you get any helpful response, could you
forward me a copy? Thank you.
P.S. Could anyone who has any comments or suggestions on this issue also
forward the response to ccp4bb? Thank you in advance.
Best,
Joe
On 7/16/07, bputcha <[EMAIL PROTECTED]>
Hi,
I am trying to crystallize a protein-DNA complex. I purify the protein finally
using gel filtration. I purchase
single stranded complementary oligos (desalting from idtdna.com), mix them up
and make DNA duplex by
heating to 95 degree C and cooling to room temperature. I mix protein and DNA,
