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
particul
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.
Yo
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,
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 of
