Help recovering 200bp DNA fragment

"Marianne Leverone ", BIO leverone at CHUMA.CAS.USF.EDU
Tue Aug 8 12:21:53 EST 1995


Harshinee:  Try something to increase ppt. of the DNA such as glycogen, a 
20 ug/ml soln. helps.  Add 1 uL per 50 uL of DNA soln.  Marianne, USF, 
Tampa, FL

On Fri, 4 Aug 1995, Harshinee Wijesinghe wrote:

>  
> I have been trying to elute and recover a 200bp insert (cut from a plasmid)
> from agarose gel without much success. I definitely know that the DNA came
> out of the gel as I monitored it under UV light and subsequently stained
> the gel slice too. The protocol I used is as follows: After elution into
> 1.5 ml of TE buffer in dialysis bags I extracted with butanol and ether
> before precipitating with ethanol. Lastly I used 0.01 MgCl2 with ethanol
> at -70 degrees C overnight and spun for 30 min. After resuspension in 15 ul
> of TE buffer and running a gel I find that although the insert is there the
> band is very faint (i.e., not much has been recovered).
>  
> The problem does not seem to be with the elution but with the recovery
> with ethanol. I would be very grateful for any suggestions for recovering
> small (e.g. 200bp) fragments of DNA. Many thanks in advance.
>  
>  
> Harshinee Wijesinghe
> vhwbc at cunyvm.cuny.edu
>  
> 
> 



More information about the Methods mailing list