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

Paul N Hengen pnh at cockleberry.ncifcrf.gov
Mon Nov 27 13:59:49 EST 1995


Alejandro Palacios F. (japalaci at cariari.ucr.ac.cr) wrote:

: 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 can't help because this means nothing to me...

: 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

Then stick with EtBr. It is not worth the cost or effort.

: am I using the wrong buffer for this dye? 

No. That should work. How old is the dye solution? If you are not seeing any
band at the expected position, and the dye is more than 3 months at -20 C, try
adding 10 ul to see the band. It could be that the dye has gone off.  I also
had this experience.

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
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