4.2.9 Protocol and Run Conditions

10Õ TBE and deionised water (Millipore) was used for the ABI377 buffer tanks. Samples were loaded in alternate wells, and run into the gels using a short (5min) prerun.

All gels were run using 1200 scans/hr run modules. Run time for 36-cm gels was 5 - 7 hours (depending on PCR fragment length), for 48-cm gels 8 - 10 hours. Prerun time was between 10 and 20 minutes. For 96 well gels, a different machine protocol (resulting in more readable channels) was used. Run temperature was 51C. Sequence analysis was done with PE Biosystems Sequence Analysis 3.4 software. Correct lane tracking was checked manually.

© 2001 Alexander Binder