One or more ingredients in the original hit (which you use to suspend the
seed crystals in) can indeed be essential, and may be all you need to get
nice crystals. In D'Arcy's original paper you will see that he needed Ca2+
to get nice crystals of one of his proteins. The same is true in Stoddards
Thanks you for the suggestions, Patrick and Matthias. I was actually
wondering if any of the components from the seeding solution actually were
important but your explanations sound more logical.
I apologize for the large attachment. I did not realize that it was so big.
Have a nice weekend !
Ma
Mahesh, this is a very interesting and slightly controversial question.
One approach is to mix together all of the crystals that you have in the
initial screen. The idea at the beginning of the project is to get as many
diverse hits as possible - you can worry about crystal size, space group
and
Dear Mahesh,
cross-seeding into other conditions often works but still the chances
are much lower than seeding the same form.
Best wishes,
Matthias
(Please desist from sending so large attachments to half the planet!)
-
Dr. Matthias Zebisch
Division of
oseph
Brock
Sent: Tuesday, September 15, 2009 7:57 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Seeding and protease questions
Hi everyone,
I have two conditions that I want to try and improve by microseeding. I have
been reading the literature on the subject and am a bit confused as to w
Hi everyone,
I have two conditions that I want to try and improve by microseeding. I have
been reading the literature on the subject and am a bit confused as to what
exactly is the "stabilizing mother liquor" that is used in the serial dilution
of the seed stock. Is this just purified protein
ee 1-877-225-2034
Regd. England 2177994, VAT Reg. GB 480 7371 36
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
HanJie_HCT Tai
Sent: 29 May 2009 21:04
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Seeding
Hi,
I set up some seeding trials (microseeding) from 1 x to 1000
Dear HengChiat Tai
(a) According to me Seed Solutions should be stored depending on the
> stability of the crystals in the drop. If your crystal goes bad quite early
> then you have to make fresh seed frequently. Its always a good habit to make
> fresh seed, everytime you do streaking.
>
> (b) Th
Hi,
I set up some seeding trials (microseeding) from 1 x to 1000x dilution.
I am just wondering
(a) How long the seed stock solutions can be kept? some said they can be kept
for up to 1 year, some said the seed stock solutions need to be freshly
prepared before use, which one is true
