A question for anyone using the Millipore/sephadex/96-well cleanup
system for 3700 sequencing.
What signal strengths are you getting?
We are test driving this system and at present are getting values
around 1200 total (A+T+C+G) with the following protocol:
Spin to pack; wash with 150ul water and spin; add 5ul water to
samples,load and spin a third time. Onto the 3700 in a resulting
volume of 12-14ul water.
We are doing 10ul reaction volumes; 0.125ug template; 5-10pmoles
primer; 1/2 BigDye (i.e. 2ul); 30 cycles.
We wonder if this is about as good as we're going to get. Or does
someone have a tweaking idea or two to further enhance.
Thanks,
Lee
Lee Eberly, RAIII phone: 319-335-6736
DNA Facility 300H, EMRB fax 319-335-6737
University of Iowa email: lee-eberly@uiowa.edu
Iowa City, IA 52240 Core WEB site http://dna-9.int-med.uiowa.edu
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