opinions on auto sequencers

Alexander Kraev kraev at bc.biol.ethz.ch
Thu Jun 20 05:05:04 EST 1996


Will Fischer wrote:

>The Licor works very well indeed, WHEN it works.  I and others from my
lab have found the chemistry for the Licor (Sequitherm kits, with dye
labelled primers) to be inordinately sensitive to template quality.
Qiagen tip DNA generally works, but the Qiawell DNA has not, in my
hands.  In contrast, the SAME TEMPLATES have sequenced very well on the
ABI.    In addition, we have had trouble with single lanes failing
(i.e., one tube out of four gives no bands at all).  This is
particularly frustrating.  One final gripe about the Licor:  the
software (OS/2 based) is not very sophisticated.  Contig assembly is
painful.  The difference between Sequencher and the Licor software is
like day and night.<

I had a very different experiecne with the LICOR: when I first tried it
(with Thermosequenase kit and dye-labelled primer), there was no 
difference in quality between Qiawell, Qiaqen 100 ug tip and 
LiCl-boiling miniprep - everything went past a kilobase! Actually, using
Thermosequenase kit with a labelled primer is not particularly difficult
either, you mix primer with template and distribute into the tubes with
"A-reagent", "G-reagent" etc and cycle. I know you cannot beat 1-tube
reaction for simplicity, but lengthy discussions in this newsgroup about
multicoloured blobs somewhat mar that picture, don't they? An ABI is 
nice when it works...


Alexander Kraev, PhD                     Internet:    
kraev at bc.biol.ethz.ch
> Lab.of Biochemistry III                            Phone:   0041-1-632-31-47
> Swiss Federal Inst. Of Technology           FAX:      0041-1-632-12-13
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