Margarita,
I have actually used Betaine, but with hit-and-miss results. On some
templates, it has worked well, and others it has not made any affect.
I have tried the Sequence Rx Enhancer system from Life Technologies. They
have 7 different buffers that you can add to your sequencing reaction, the
"F" buffer is supposed to be for GC-rich templates. This I found to be the
case for a difficult template I was trying to sequence. What I did notice,
was that I tried 4 different primers in the difficult area, and only one
primer worked really well with the "F" buffer. The other buffers worked OK
with some of the other buffers. I got ~550bp with "F" buffer with the one
primer. The other buffers with the other primers got ~350bp on average.
Overall, I got through the difficult area.
Hope this info. is helpful.
Dina
Dina Leviten
ICOS, Corp.
> ----------
> From: Margarita Almeida
> Sent: Thursday, May 25, 2000 5:56 AM
> To: Recipients of ABRF List
> Subject: Sequencing difficult templates
>
> Hello. I am having difficulty sequencing GC rich templates. I have a 373
> with XL upgrade and use Big Terminator chemistry. I have heard of a
> product
> called Betaine. Has anyone used it or does anyone have any other
> suggestions?
>
> Thanks,
> Margarita Almeida
> University of Miami School of Medicine
> Department of Biochemistry and Molecular Biology
> DNA Core Facility
>
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