SOS Call to all experinced "Sequencers"

Pierre-Alain Menoud menoud at cmu.unige.ch
Tue Apr 2 11:56:16 EST 1996


In article <199603281727.BAA03464 at relay3.jaring.my>,
   spyong at pc.jaring.my wrote:
>Hello,,
>
>The only sequencing I've ever done is through the text book. However, it
>seems like now is the time to start sequencing. I have three PCR products
>and need to sequence them before I work further only to find that they are
>junk. Someone suggested end-labbeling, others say double strand sequencing,
>some say cycle sequencing....some say stick to 'Sequanase' (whatever that
>means). Do I have to subclone into M13?
>
>Please,I know sequencing is not easy and may be take ages, but I just need


Hello Stephanie,

What I usually do is the direct sequencing of my PCR product using the "fmol 
sequencing kit" of Promega. Its relatively cheap and very easy to perform. You 
just need to kinase your primer. Everything is within the kit.
If my PCR product is not pure enough, I purify it on agarose gel and extract 
the fragment with Qiaex or Geneclean stuff.

Good luck

Pierre-Alain
menoud at cmu.unige.ch



More information about the Methods mailing list