The concentration of oligonucleotide seems much lower than you will ultimately
need for biophysical work. Annealing two strands is a bimolecular reaction and
it will be driven by mass action more to completion at higher total
oligonucleotide concentrations. I also agree with the posts that hav
Le Mardi 7 Juillet 2015 14:03 CEST, Almudena Ponce Salvatierra
a écrit:
Hello
Do you have full complementarity of DNA and RNA ?
Does the difference in lenght (17 nt/19 nt) mean that you have a bulge on the
DNA when the duplex is formed ?
In short, what is the Tm of this duplex at these strand
Hi Weifei,
so... what I do is to mix them in a 1:1ish ratio, then I check that ratio
on the HPLC in case I can adjust it to 1:1. I.e. Instead of mixing 30 nmol
of RNA with 30 of DNA if I want a 1:1 ratio complex, I mix 30 of RNA and 25
of DNA. I calculate their molar extinction coefficient (epsilo
board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of ChenWeiFei
Sent: 03 July 2015 16:14
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] DNA RNA annealing problem
Dear all,
I want to get a complex of DNA-RNA-protein. But I have a big problem of
annealing DNA-RNA.
The length of DNA is 19nt and RNA is 17nt
Hi Weifei,
We use SSC buffer for annealing oligos (we use DNA). That works pretty fine
with us. Give a try. Composition is in below URL
http://www.bioprotocols.info/reagent_and_buffer_recipes/20x-SSC.php
Good Luck,
Venkatareddy.
On Fri, Jul 3, 2015 at 8:44 PM, ChenWeiFei wrote:
> Dear all,
>
Dear all,
I want to get a complex of DNA-RNA-protein. But I have a big problem of
annealing DNA-RNA.
The length of DNA is 19nt and RNA is 17nt.
Annealing protocol:
2uM DNA
2uM RNA
10mM Tris-Hcl
100mMNaCl
1mMEDTA
Heated to 95 for 5min, cooling down slowly for nearly 2h to room temperature.
I can
