Dye-Deoxy Cycle Sequencing-ramp time?

Thomas Newman 22313tcn at ibm.cl.msu.edu
Mon Jan 30 17:51:39 EST 1995


In article
<Pine.OSF.3.91a.950127195746.12261B-100000 at saul5.u.washington.edu>, James
Gray <jp at u.washington.edu> wrote:

> 	Does anyone know how important ramp time REALLY is in Taq 
> Dye-Deoxy cycle sequencing?  I am using an MJ Research Minicycler using 
> the 0.2ml tubes and the default ramp times on the cycler.  This machine 
> has a noticeably faster ramp time than the recommended Perkin-Elmer 
> machine (1C/sec).  I find that my reads fall off at about 90 bases and 
> the background is high.
> 

We just ran some DNA samples (0.2ug/um promega wizard prepped) on the new
MJR DNA Engine, with 3C/sec ramp times and the results were fine.  If the
initial peaks from the sequencer are very strong, you may be starting with
too much DNA in the sample which results in early garbage. 

Tom Newman
MSU-DOE Plant Research Lab
Sequencing Facility 



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