One problem may be the thickness of your gel. We recommend that you
use only 3-4mm thick gels for the normal staining time of 20-30
minutes. Also, our protocol states a pH range of 7.0-8.0... dilutions
made in TE or Water are fine. Do you use glyoxyl in your RNA gels? If
so, before staining with SYBR you might want to try immersing the gel
in 0.5M ammonium acetate buffer for 45-60 minutes. This will aid in
the staining, as SYBR and EtBr do not stain glyoxylated RNA well.
Please let me know if you have other questions. You may also call us
at 800-521-0390
Steven Gerhardt
FMC BioProducts
Technical Service
______________________________ Reply Separator
_________________________________ Subject: SYBR II problems
Author: mjmgmatos at gemini.ci.uc.pt (Manuel J. Matos) at Internet Date:
10/25/96 4:34 PM
I am having problems with SYBR Green II (FMC) for RNA staining. After
running the RNA agarose gels, I immerse them in 10 mM sodium phosphate
buffer, pH 6.8 and a 1:5,000 SYBR Green II (as indicated by FMC).
Bands become clearly visible over a transilluminator only after 4
hours of incubation, instead of the 30 minutes mentioned by FMC. Hints
anyone?
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Manuel J.
Matos (Manolo) -- mjmgmatos at gemini.ci.uc.pt * * Departamento de
Bioquimica * Universidade de Coimbra * * Aptd. 3126 * P - 3000
Coimbra * P O R T U G A L * * tlf. +351-39-22174 *Macintosh user*
fax +351-39-33827 * * * * * * * * * * * * * * * * * * * * * * * * * *
* * * *
