faint bands

Ma Sheila M. De Jesus mshmdeje at USTCC.ust.edu.ph
Sun Mar 5 18:33:24 EST 2000





On Fri, 3 Mar 2000, Anne Gillen wrote:

> I'm confused, did you get a band of the correct size?  I'll assume you
> have the correct product.

Yes, I got the band of teh correct size.


> 
> What staining method are you using?  When you said you ran the gel a
> second time do you mean you put it back in the gel box and continued to
> run it, or did you start from the beginning and run the PCR samples in a
> new gel.  Are you size markers faint? Perhaps you need a bit more ethidium
> bromide.  Remember that ethidium bromide moves in the opposite direction
> as the DNA during electrophoresis. 
> 
I used EtBr staining......my markers were not faint.....and yes, i ran the
samples in a new gel.  

>  You can always load more product to get a brighter band, but I assume
> you know this already.  If it really is a PCR problem, then I suggest
> optimizing your reaction. Especially the annealing temperature and the
> magnesium chloride concentration.  

Thanks to all who cared to answer.....

> 
> Anne
> 
> 
> On 3 Mar 2000, Ma
> Sheila M. De Jesus wrote:
> 
> > 
> > Hello,
> > 
> > I see faint bands in my gel (from PCR) but they are the bands of interest
> > (correct size).  What does it mean?  How can I make it look brighter?
> > 
> > Second question is how come, on my second time i run the gel, i couldn't
> > see the bands anymore.
> > 
> > Third, what is the best diluent for dNTPs.
> > 
> > Thanks in advance.
> > 
> > 
> > 
> > ***************************************************
> > MA. SHEILA M. DE JESUS				 **
> > Dept. of Biological Sciences			 **
> > College of Science				 **
> > University of Santo Tomas			 **
> > Espana, Manila					 **
> > 1008 Philippines				 **
> > 						 **
> > Fax: +63(2)-740-9730				 **
> > Tel.: +63(2)-731-3001 to 12 ext. 8297		 **
> >       +63(2)-731-8301 to 17 ext. 8297		 **
> > 						 **
> > email: (home) boston at quickweb.com.ph		 **
> >        (work) mshmdeje at ustcc.ust.edu.ph		 **
> > ***************************************************
> > 
> > 
> > ---
> > 
> > 
> 
> *******************************************************************************
>     Anne M. Gillen                           E-mail: amgillen at ucdavis.edu
>     Pomology Department                      Phone:  (530) 752-1462
>     University of California at Davis       
> *******************************************************************************
> 
> 

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