SYBR Green I Dye for agarose gel eletroph. of DNA?

Tracey Ann Ristow Lozano tar205 at nwu.edu
Tue Nov 28 11:54:45 EST 1995


In article <Pine.A32.3.91.951125154854.79008B-100000 at cariari.ucr.ac.cr>, 
japalaci at cariari.ucr.ac.cr says...
>
>
>Hi bionetter,
>
>  I am using a dye named SYBR Green I with no good results.
>  I have heard that this dye make a better contrast in the UV light and 
>then better sensitivity in nucleic acids agarose gel electroforresis  but 
>a have amorphus (abnormal morphology) of the bands.
>  I do the electroforetic run at:  100volt/300mA/50W and 1.2 Sigma 
>agarose for a PCR product of 207 bp.  Using TBE 0.5X
>  I use 1.5 microliter of green per 100ml of agarose
>
>This conditions have work for a long time using Etidium bromide but not 
>with this dye.  
>The test using higher quantitiest increase the problem
>
>       am I using the wrong buffer for this dye? 
>
>Thanks is advance for your sugestions,
>
>
>-------------------------------------------------------------------
>Alejandro Palacios           /  International Center for Medical  
>japalaci at cariari.ucr.ac.cr   /  Research and Training (LSU-ICMRT) 
>------------------------------------------------------------------
>


We have always had bad luck adding SYBR Green directly to the gel. We stain 
the gel after instead, with much better results.  We have to make the stain 
up at 2x the recommended strength though, and this gets kinda costly.  But 
the pictures we get are much better than with Ethidium bromide, and you 
don't have to de-stain it.
-- 
Tracey Ann Ristow Lozano
Northwestern University, Evanston, IL.   USA
tar205 at nwu.edu




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