Hi,
Sorry for the non-crystallography question, does anyone know how to produce
milligram quantities of single stranded DNA? Thanks.
Best wishes,
Reza
Reza Khayat, PhD
Assistant Professor
Department of Chemistry
City College of New York
85 Saint Nicholas Terrace, CDI 12318
New York, NY 10031
ht
1) Commercially made oligonucleotides
2) M13 rolling circle replication (how people used to make ssDNA for sanger
sequencing).
———
Dr Antony W Oliver
Senior Research Fellow
CR-UK DNA Repair Enzymes Group
Genome Damage and Stability Centre
Science Park Road
University of Sussex
Fal
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
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,
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
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
