User guide

22
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter2 Perform PCR
Treated paper substrates: prepare reactions
2
3. Prepare reagents. Thaw the Master Mix and the Primer Set, then vortex 3 seconds
and centrifuge briefly before opening the tubes or bottles.
IMPORTANT! Thawing is required only during first use of the kit. After first use,
reagents are stored at 2 to 8 °C and, therefore, do not require subsequent thawing.
Do not refreeze the reagents.
4. Pipet the required volumes of components into an appropriately sized
polypropylene tube.
5. Vortex the reaction mix for 3 seconds, then centrifuge briefly.
6. Dispense 15 µL of the reaction mix into each reaction well of a MicroAmp
®
Optical 96-Well Reaction Plate.
7. Seal the plate with MicroAmp
®
Clear Adhesive Film or MicroAmp
®
Optical
Adhesive Film.
IMPORTANT! We recommend adhesive film for plate sealing to provide a
consistent seal across all wells and prevent evaporation. Do not use caps, which
may not provide a consistent seal across all wells.
IMPORTANT! If using the 9700 thermal cycler with silver or gold-plated silver
block, place a MicroAmp
®
compression pad (Part no. 4312639) on top of the plate
to additionally prevent evaporation during thermal cycling. The Veriti
®
Thermal
Cycler does not require a compression pad.
8. Centrifuge the plate at 3000 rpm for about 20 seconds in a tabletop centrifuge
with plate holders.
9. Amplify the samples in a Veriti
®
96-well Thermal Cycler or PCR System 9700
with the silver or gold-plated silver 96-well block as described in “Perform PCR”
on page 28.
IMPORTANT! This kit is not validated for use with the GeneAmp PCR System
9700 with the aluminium 96-well block. Use of this thermal cycling platform may
adversely affect performance of this kit.