User guide

21
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter2 Perform PCR
Treated paper substrates: prepare reactions
2
Prepare low-TE
buffer
You can prepare the buffer as described below or order it from Teknova (Cat # T0223).
To prepare low-TE buffer:
1. Mix together:
• 10 mL of 1 M Tris-HCl, pH 8.0
• 0.2 mL of 0.5 M EDTA, pH 8.0
• 990 mL glass-distilled or deionized water
Note: Adjust the volumes accordingly for specific needs.
2. Aliquot and autoclave the solutions.
3. Store at room temperature.
Prepare the
reactions
1. Add samples to the reaction plate:
2. Calculate the volume of each component needed to prepare the reactions, using
the table below.
Note: Include additional reactions in your calculations to provide excess volume
for the loss that occurs during reagent transfers.
IMPORTANT! This kit has been optimized for a 15-µL PCR reaction volume to
overcome the PCR inhibition expected when amplifying unpurified samples.
Using a lower PCR reaction volume may reduce the ability of the kit chemistry to
generate full STR profiles.
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 • 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
IMPORTANT! Do not add a blank disc to the positive control well.
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.
Reaction component Volume per reaction
Master Mix 6.0 µL
Primer Set 6.0 µL
Low TE buffer 3.0 µL