abi 373 questions

Scottie Scottie
Tue Apr 1 19:38:23 EST 1997


Hi

        We have an antique 373A using 24 cm WTR plates and a 24 lane
format.  We routinely use half reactions with no noticeable loss of
sequence accuracy. Some students have trouble with the 10 ul volume - so
they still use 20 ul total volume but only use half the premix (4ul) and
even this seems to work. There will be a webpage version of the ABRF DNA
Sequencing  Research Committee Report released vey soon in which many
people used half reactions (but not necessarily on the 373) and at least
one person has reported using 1/4 reactions with success.

        As in any situation - you just have to try it for yourself and see
if it works - but it is cetainly feasible.   Hope this helps.  Scottie

>does anyone know if half reactions or smaller(10 microliter total volume
>sequencing reaction or less) can be used per lane on ABI 373  machines
>with decent reads (400-500  bases)?
>if yes are there any tricks?
>i also have the same question about the ABI 373 XL machines using the 64
>well format.

Scottie Adams aka Pamela Scott Adams
Coordinator
Molecular Biology Core Facility
W. Alton Jones Cell Science Center
10 Old Barn Road
Lake Placid, NY    12946
Phone:  518-523-1276
Fax:    518-523-1849
Email: scottie at cell-science.org
http://www.cell-science.org/mbcf





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