SYBR II Problems

STEVEN GERHARDT steven_gerhardt at FMC.COM
Mon Oct 28 13:58:45 EST 1996


     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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