User guide

27
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter2 Perform PCR
Swab substrates: prepare reactions
2
6. Dispense 12 µL of the reaction mix into each reaction well of a MicroAmp
®
Optical 96-Well Reaction Plate. The final volume in each well is 15 µL (reaction
mix plus Prep-n-Go™ Buffer and sample lysate or positive control).
7. Add samples to the reaction plate:
The final volume in each well is 15 µL (reaction mix plus Prep-n-Go
™
Buffer and
sample lysate or positive control).
8. 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.
9. Vortex the reaction mix at medium speed for 3 seconds.
10. Centrifuge the plate at 3000 rpm for about 20 seconds in a tabletop centrifuge
with plate holders.
11. 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.
Store the sample
lysate
Cap the sample lysate storage tubes or seal the sample lysate storage plate with
MicroAmp
®
Clear Adhesive Film.
Store the sample lysate as needed:
Well(s)
Add the following to wells of a MicroAmp
®
Optical
96-Well Reaction Plate...
Test samples 3 µL of sample lysate
Positive control • For 25 and 26 cycles 3 µL of DNA Control 007
• For 27 cycles 2 µL of DNA Control 007
• For 28 cycles 1 µL of DNA Control 007
Note: The volumes of positive control are suggested amounts and may be adjusted
if peak heights are too high or too low for your optimized cycle number.
If you are storing the sample lysate... Then place at...
<2 weeks 2 to 8°C
>2 weeks –15 to –25°C