High Performance Gel Electrophoresis (HPGE)

Gel of the Month 

Author(s):
Anonymous (Upon request)
Institution:
Industry [Research Facility, Biotech, CA, USA.]
Experimental:
Temperature gradient profiling of site mutations on PCR
amplified dsDNA fragments: Real-time analysis of melting/
annealing conditions, using timed temperature gradient
electrophoresis fluorescence profiles.
HPGE File:
101298NA6.dat [Experiment date: 12/10/98]
Comments:
Gel: 5%T, 5%C (BIS) Polyacrylamide gels [3mm, clear trays],
2M Urea, 30mM TAE (pH 8.30).
Samples: 15µl load / lane (~30ng total dsDNA)
Temperature ramping: from 40-65°C up in 1°C steps,
then down to 55°C (1°/ 6 min.).
Separation conditions: 15 V/cm constant; continuous
TAE buffer system.
Scanning: every 6 min., synchronized with temperature ramp.
 
March 1999
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