more 96 lane comments
Margaret Robertson (mar@howard.genetics.utah.edu)
Mon, 24 Aug 1998 10:29:19 -0600 (MDT)
We have now tried the 48cm plates on the 96 lane upgrade with good
success for longer reads:
We use Rapid Ranger (4.5%) running the 48e-1200 module at 2200 volts.
This gives us 750 bases over a 10 hour run. We reduced the voltage
slightly to prevent getting red rain. the pGEM control reads base 2 after
the primer, first N at 640, with 4 Ns total between 600 and 700 bases.
Inserting the 96 lane comb to get good even lane formation, seems to be
critical for good resolution. Our 96 lane combs are very tight to insert
even though we clamp with loose clamps, not the ABI clamps sent with the
upgrade. We're loading 1ul out of 4ul, = 150 ng DNA, using a new
expandable tray and new "prizm" strips for 10.8 mm spacing. See the
Sooner Scientific web page at www.soonersci.com or EmbiTec 1-800-255-1777.
My tecnnicians find the pipette very reliable and much easier to use than
the Kloen adjustable pipette. There is no washing required as the strips
are disposable and requires much less repetitive movements.96 lane Gels
can be loaded in 8 minutes with very little chance os sample mix up.
Margaret Robertson, Director, DNA Sequencing, University of Utah