Getting a 373 stretch. Any tips?

Kevin Clark kclark at immsvr.jr2.ox.ac.uk
Tue Jul 9 05:50:28 EST 1996


We are getting a stretch upgrade to our 373 in a couple of weeks and I
would be grateful for any tips users have.

I currently run 85% M13 clones (PEG/phenol preps) and 15% pcr products
(PEG/NaOAc preps) all with ABI Taq FS terminators. If possible I do not
want to take on a new prepping method to get long reads on the 48cm wtr
plates as the current ones are high quality, quick, easy, reliable and
cheap. Will this quality of prep be OK for long reads? Do I need to
change the sequencing reaction conditions to favour longer products?

Also I know the gel will have to be 4.75% (4.25%?). I currently use
sequagel-6 which is great. Do you recommend any particular ready mix
for the long gels?

Finally can anyone give me some typical read lengths for the stretch
machine.

Thanks in advance

Kevin

Kevin Clark                    |
kclark at immsvr.jr2.ox.ac.uk     | If you havn't seen God, you're not
Institute of Molecular Medicine| driving fast enough.
Oxford,UK.                     |      




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