User guide

33
GlobalFiler
™
Express PCR Amplification Kit User Guide
Section3.1 3500/3500xL instruments
Prepare samples for electrophoresis on the 3500/3500xL instruments
3
3500/3500 xL Instruments
Prepare samples for electrophoresis on the 3500/3500xL
instruments
Prepare the samples for electrophoresis immediately before loading.
1. Calculate the volume of Hi-Di
™
Formamide and GeneScan
™
600 LIZ
®
Size
Standard v2.0 needed to prepare the samples:
Note: Include additional samples in your calculations to provide excess volume
for the loss that occurs during reagent transfers.
IMPORTANT! The volume of size standard indicated in the table is a suggested
amount. Determine the appropriate amount of size standard based on your
experiments and results.
2. Pipet the required volumes of components into an appropriately sized
polypropylene tube.
3. Vortex the tube, then centrifuge briefly.
4. Into each well of a MicroAmp
®
Optical 96-Well Reaction Plate, add:
• 10 µL of the formamide:size standard mixture
• 1 µL of PCR product or Allelic Ladder
Note: For blank wells, add 10 µL of Hi-Di
™
Formamide.
5. Seal the reaction plate with appropriate septa, then briefly vortex and centrifuge
the plate to ensure that the contents of each well are mixed and collected at the
bottom.
6. Heat the reaction plate in a thermal cycler for 3 minutes at 95°C.
7. Immediately place the plate on ice for 3 minutes.
8. Place the sample tray on the autosampler.
9. Start the electrophoresis run.
Reagent Volume per reaction
GeneScan
™
600 LIZ
®
Size Standard v2.0 0.5 µL
Hi-Di
™
Formamide 9.5 µL