Dear all,
Recently,i purchase Oligonucleotide from company.the sequence is
TTGCGTAC GCAC GTACGC .i want to perform self-annealing process to form
the following second structure .
5' TTGCGTACGC
||| |||]
3' CGCATGCA
i hope that most
Just like you would for PCR:
- Dissolve the oligonucleotide in water
- Heat it to above meltung temperature, pick 95degree C if you want to be on the
save side
- leave it to cool down.
Is this what you are looking for?
Tim
On Thu, Mar 17, 2011 at 07:23:07PM +0800, dengzq1987 wrote:
Dear all,
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I would bring up the DNA in TM buffer (10 mM Tris, 5 mM MgCl2) or similar
and anneal under dilute conditions to favor hairpin formation over dsDNA.
Fast cooling will also favor hairpin formation, so you may try heating to
95° and then cooling on ice, or using a short gradient on a thermocycler.
Has anyone looked at the kinetics of DNA annealing. Especially for
such short fragments I expect hairpin formation times to be on the
order of pico to nano seconds. Of course it doesn't hurt to
slow-cool but I wouldn't be too paranoid about it. Moreover, in this
I seem to have noticed that (i)mosflm can index on one image, or two
images that are not related by 90 degrees. also, cell refinement
sometimes splits up a set of frames into maybe 3 or four segments of
e.g. a few degrees each. and of course, I can set it based on frames
90 degrees apart.
reason
afaik Mosflm will index on whatever spots you have told it to find and not
discard, be they from 1 image, 2, or 10 or more images at any relative angle or
oscillation range. Or it won't successfully index, depending on the difficulty
of the case (several lattices, spot overlap etc...).
Indexing
We have a very low mileage rotating anode (Rigaku RU-H3RHB 18KW Cu anode, 0.3mm
focus) which has been retired for a while now following arrival of an Xcalibur.
There are also lots of spares, some new. And a table top enclosure. Free to
anyone
who is able to come and get it.
Trevor Greenhough
--
Dear all,
Thank you very much for all of information about domain determination! I
think the limited proteolysis is a good choice for the domain determination
as we have no information about a protein.
However, if we do have a domain information by the bioinformatics, how can
we truncate the
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yes limited proteolysis is the best choice for the domain determination of
unknown protein from our lab some people did this. after doing
limited proteolysis just sequence digested part N terminally or the C
terminally and find out the region from where its getting digested.
side by side u can
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