Cycle sequencing

Bill Burnett bbur at wpo.nerc.ac.uk
Tue Oct 17 14:48:03 EST 1995


In article <460733$d3s at whitbeck.ncl.ac.uk> n4522701 <R.Guy.Reeves at ncl.ac.uk> writes:
>From: n4522701 <R.Guy.Reeves at ncl.ac.uk>
>Subject: Cycle sequencing
>Date: 17 Oct 1995 12:20:51 GMT

>I am thinking of trying out Cycle  sequencing of PCR products.
>Does any body have advice or tips on what is an effective kit,
>is cycle sequencing tricky to do???  How little DNA do I need
> to be successfl.


>Thanks

>Guy Reeves 

>Dpt of Biological & Nutritional Sciences
>Agriculture building
>Newcastle University
>Newcastle upon Tyne
>NE1 7RU
>Fax (0) 191 222 8684
>R.Guy.Reeves at ncl.ac.uk

I used an automated system and Applied Biosystems PRISM DyeDeoxy Cycle 
sequencing kit.
It's a doddle, easier to set up than a PCR BUT if I was going to do it again I 
would clone first.  Heck, I AM  going to do it again :-).
You need very little template, 100-200 ng springs to mind.

Don't know about manual systems, I imagine the only complication is doing it 
in four tubes in stead of one?  Correct me someone if I'm wrong.

Good luck!  And give my love to the Bacchus, the Red House, the Duke of York...
*sigh* days gone by....

Bill

 



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