Annealing Oligos

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Tue Feb 22 15:04:48 EST 2000


Andy, 
use 10ul of 1uM oligo in 1x hybridization buffer 
(10x is 200mM trisHCl pH7.5, 100mM MgCl2, 350mM NaCl)
place in thermocycler
heat 2min @ 94degC, cool down slowly to room temp using a gradient 
for say 60 min. you may quickly cool to 70 to 75 deg befor you run 
the gradient, since your oligo is not long. then use an aliquot (e.g. 
1 ul) for ligation.

Wolfgang

> From:          Andy Scotter <pcxajs at unix.ccc.nottingham.ac.uk>
> Subject:       Annealing Oligos
> Date:          Tue, 22 Feb 2000 16:36:37 +0000
> Organization:  ACS, The University of Nottingham
> To:            methods at hgmp.mrc.ac.uk

>  I've got a palindromic 31 base oligo that I want to anneal with
> itself so I can blunt clone it into pT7Blue-2.
>  Can anyone suggest reasonable conditions and amounts to carry out
> the annealing?  I guess it shouldn't be too hard as the oligos
> should stick like glue.  After the annealing step I'll be purifying
> the product before the end conversion step and ligation.
> 
> Cheers,
> 
> Andy
> 
> ********************************************************************
> **********
> 
> Andy Scotter
> Department of Chemistry
> University of Nottingham
> University Park
> Nottingham
> NG7 2RD
> 0115 9514193
> pcxajs at unix.ccc.nottingham.ac.uk
> 
> ********************************************************************
> **********
> 
> 
-----
This message is encrypted. Use your brain to decode it.
-----
Dr. Wolfgang Schechinger, Dept. of Pathobiochemistry
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de 
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-----
usual disclaimers apply 
-----
---




More information about the Methods mailing list