User guide

41
GlobalFiler
™
Express PCR Amplification Kit User Guide
Section3.3 3730 instrument
Prepare samples for electrophoresis on the 3730 instrument
3
3130/3130xl Instruments
Perform spectral
calibration
Perform a spectral calibration using the DS-36 Matrix Standard (J6 Dye Set)
(Part no. 4425042). The following figure is an example of a passing 6-dye spectral
calibration.
Prepare samples for electrophoresis on the 3730 instrument
Prepare the samples for electrophoresis immediately before loading.
1. Calculate the volume of Hi-Di
™
Formamide and size standard 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:
• 9 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.
Reagent Volume per reaction
GeneScan
™
600 LIZ
®
Size Standard v2.0 0.5 µL
Hi-Di
™
Formamide 9.5 µL