Dye Cleanup and Big Dye ver. 3?

Dr Mick Jones m.d.jones at ic.ac.uk
Wed Apr 24 11:41:47 EST 2002


Hi

I=B9ve just taken over (well for ~6 months plus) running a Core DNA sequenc=
ing
lab, which has an ABI 3700 CE sequencer.

I guess these questions have probably been asked before, but hopefully they
can stand re-asking and re-evaluating.

1. Currently we use a Genetix gel filtration plate (centrifugation) to clea=
n
up our Dye terminator reactions before loading on the sequencer.  Do people
feel this is a good method (we have to consider that we receive variable
quality DNA preps for sequencing, so we have to have a robust method)?
Also, do people just use iso-propanol pption?  Or EtOH pptions?
2. We are using version 2 of the Big Dyes, as we don=B9t seem to get better
results with version 3 (although we haven=B9t pushed it yet).  Would people
recommend going for ver. 3?  and have you any protocol tips?

Many thanks in advance, and its good to be back in a sequencing lab after a
period of mainly teaching MSc students.

Mick


Michael D Jones, BSc, PhD
Head Genomics Core Laboratory

MRC Clinical Sciences Centre
Room 207, L Block
Faculty of Medicine
Imperial College of Science, Technology and Medicine
Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN
Tel:  020-8383-1643 (MRC GCL), 020-8383-3328 (IS);  Fax:  020-8383-8577 (MR=
C
GCL)
Email:  mick.jones at csc.mrc.ac.uk





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