User guide

23
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter2 Perform PCR
Untreated paper substrates: prepare reactions
2
Untreated paper substrates: prepare reactions
Sample prep
guidelines
• If you are processing a blood sample collected on untreated paper substrate,
make the punch as close as possible to the center of the sample to ensure optimum
peak intensity. Increasing the size of the punch may cause inhibition during PCR
amplification.
• If you are using a Bode Buccal DNA Collector
™
,
make the punch as close as possible to the tip of the
DNA collector to ensure optimum peak intensity.
Increasing the size of the punch may cause
inhibition during PCR amplification.
• For manual punching: Place the tip of a 1.2 mm
Harris Micro-Punch on the card, hold the barrel of
the Harris Micro-Punch (do not touch the plunger),
gently press and twist 1/4-turn, then eject the punch
in to the appropriate well on the reaction plate.
• For automated punching: Please refer to the User Guide of your automated or
semi-automated disc punch instrument for proper guidance.
Prepare the
reactions
1. Add Prep-n-Go
™
Buffer (Part no. 4467079) to the reaction plate:
2. Add samples to the reaction plate:
3. Centrifuge the plate to ensure the punches are immersed in the Prep-n-Go
™
Buffer.
Take punch
as close to
the tip as
possible
Bode Buccal
DNA
Collector
™
To these well(s) of a
MicroAmp
®
Optical
96-Well Reaction Plate...
Add:
Negative control 3 µL of Prep-n-Go
™
Buffer
Test samples 3 µL of Prep-n-Go
™
Buffer
Positive control • For 25 and 26 cycles 0 µL of Prep-n-Go
™
Buffer
• For 27 cycles 1 µL of Prep-n-Go
™
Buffer
• For 28 cycles 2 µL of Prep-n-Go
™
Buffer
To these well(s) of a
MicroAmp
®
Optical
96-Well Reaction Plate...
Add:
Negative control 1.2 mm blank disc
Test samples 1.2 mm sample disc
Positive control
IMPORTANT! Do not add a
blank disc to the positive
control well.
• 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.