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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 *
* * * * * * * * * * * * * * * * * * * * * * * * * * * * *
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From: mjmgmatos at gemini.ci.uc.pt (Manuel J. Matos)
Subject: SYBR II problems
Date: Fri, 25 Oct 1996 16:34:27 -0100
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