AmpFlSTR Identifiler Amplification using TC 9700
____ 1. Bring each sample to be amplified to 10 L with TE-4, if necessary.
For the positive amplification control, use 10 L (1 ng) of Control DNA 9947A from kit.
For the negative amplification control, use 10 L TE-4.
____ Prepare the 10 L sample to be amplified in a Thin -Walled GeneAmp tube or 96-well amplification plate.
OR
____ Prepare the 10 L sample to be amplified in a transfer tube.
____ 2.
Determine the number of samples to be amplified: ____________ (Kit lot # ______________________)
# of samples x 10.5 L = ___________ L of PCR Reaction Mix
# of samples x 0.5 L = ___________ L of AmpliTaq Gold DNA Polymerase
# of samples x 5.5 L = ___________ L of AmpFlSTR Identifiler Primer Set
____ 3.
____ Deliver 15L of the above master mix into each sample tube or into the corresponding wells of a 96-well
amplification plate.
OR
____ Deliver 15 L of the above master mix into each Thin-Walled GeneAmp tube, or a 96-well amplification
plate. Add 10 L of sample to each tube or well.
____ 4.
Choose User identi and Method Identifiler. Thermal cycler #: _______________.
Verify parameters:
____ 5.
Taq Activation @ 95°C, 11 min;
28 cycles:
Denature @ 94°C, 1 min,
Anneal @ 59°C, 1 min,
Extend @ 72°C, 1 min;
Final Extension @ 60°C, 60 min;
Hold at 5°C.
Start Method Identifiler.
Case #(s): ___________________________
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Date/Analyst: ___________________________
Page: ___________________________
Issue Date: 08/01/08, Revision 0