Hello Gloria,
I usually let Coot display the nearest symmetry molecules (as CA) and save
those that fit: Option Save Symmetry Coordinates... and pick the
molecule to save.
Best regards, Daniel
--
Daniel Schlieper email: daniel.schlie...@tuxomania.net
Biochemische
can any one help me in suggesting that what mistake i have did in my mutagenic
pcr . actually my problem is my primer annealing temperature is 81degree. im
using phusion pol enzyme. i have made many trial, i.e., made annealing at 68
and then followed 2 step pcr method and then added 1micro lit
Dear Crystallographers,
Thank you for your responses. The density map levels were 0.11e/A^3 (3.00 A)
for both images with and without TLS refinement. When I superposed the
deposited structure I could see the extra density was due to water moleucles
that were seen in the higher resolution
Hello Fellow PHENIX users (and others!)
I had been using PHENIX to generate iterative build composite OMIT maps. I was
able to perform these runs with automated settings for two of my data. With the
third one (1.2 A structure) I get an error. I am pasting a part of the message
below:
At the Buchmann Institute of Molecular Life Sciences (former Frankfurt
Institute of Molecular Life Sciences; Goethe University
Frankfurt/Main) there is an opening for a Post Doctoral Position (E13
TV-G-U, fulltime). The position is available from April 1, 2012, and
the initial appointment
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Hash: SHA1
Dear Arun,
it's been a while for my last PCR, so take my comment with caution:
33bases seems very long for a primer especially if it is only for a
point mutation. Could you trim it down to 20-25
bases? I would cut at the downstream side.
Tim
On
Dear Arun,
I find stratagene method using Pfu turbo enzyme much more successful then using
phusion. If you go to their website they even have a tool to design the
'perfect' primer!
Happy cloning!
On 24/02/2012 10:54, Tim Gruene t...@shelx.uni-ac.gwdg.de wrote:
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Dear Naveed A Nadvi
I think that your results highlight the fact that modelling the
disorder/complex ordering of your crystal is relevant and in general,
that we should take care in optimizing B factor refinement as a strong
factor for model improvement.
In this sense I would not relay
Phusion requires that the primers are phosphorylated for mutagensis to work,
unlike Pfu. If you cannot phosphorylate them use Pfu as recommended by Charlotte
Dan
Does anyone know of a program that will output sigma_a and D values for
individual reflections (yes, I know they're usually calculated as bin
values)? Or, for that matter, an option to an existing program I
haven't found in the documentation yet?
Thanks,
Pete
Phusion requires that the primers are phosphorylated for mutagensis to
work, unlike Pfu. If you cannot phosphorylate them use Pfu as recommended
by Charlotte
Not really. Phusion *protocol* requires phosphorylated primers but
seemingly the only reason for that is that they needed to find a way
Hi Pete,
yes, I wrote one a while ago:
phenix.reciprocal_space_arrays
https://www.phenix-online.org/documentation/reciprocal_space_arrays.htm
it does exactly what you asked for and much much more.
Let me know if you have questions.
Pavel
On Fri, Feb 24, 2012 at 9:28 AM, Pete Meyer
Hi Pavel,
Thanks - I'd missed that one when I was checking the phenix documentation.
Also, I may be slower than usual today, but I don't see an output column
for sigma_a. Is there a relationship between alpha and beta and sigma_a?
Pete
Pavel Afonine wrote:
Hi Pete,
yes, I wrote one a
All - See the link below; there was a story on NPR Science Friday
about a photographer who took videos of ice crystals forming under
cross-polarizers. Then, they said they want listeners to send in cool
science videos - I bet there are some of you out there (not me) that
would have the equipment
Please excuse cross postings and feel free to forward.
Dr. Soares Netto was a NIH Postdoctoral Fellow in the Laboratory of
Biochemistry (NHLBI) with Dr. Stadtman from 1992-1996.
*Post-doctoral position in Biochemistry and Structural Biology with
emphasis on Antioxidant Proteins.*
*University of
Hello Arun:
Actually, I am not sure about i couldn't get my desire point mutation. You
mean you didn't get the pcr product or you could get the pcr product but there
is no mutation. If you didn't get the PCR,Just lower the annealing temperature
to 55 degree. And try extension temperature 72
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