Profiler Plus or COfiler Amplification Set-Up
Standards and Controls:
1) Reagent Blank: as specified in the extraction procedure used
2) Substrate Control: if used in the extraction process
3) QC sample: one required per case
4) Positive Amplification Control: extracted DNA of specific, known type supplied with the Profiler
Plus or COfiler Typing kit plus amplification reagents
5) Negative Amplification Control: Profiler Plus or COfiler amplification reagents, but no added DNA
1. Retrieve amplification reagents from an inventoried kit. Label the thin wall gene-amp tubes to match your samples. Kit Lot No.: ______________ Control Date:__________________
2. Prepare the Profiler Plus or COfiler Master Mix by adding the following volumes of reagents to a
1.5 ml tube:
# of samples = ____× 21.0 
L Reaction Mix = ____

L.
# of samples = ____× 11.0 
L Profiler Plus or COfiler Primer Set = ____

L.
# of samples = ____× 1.0 
L AmpliTAQ Gold DNA Polymerase = ____

L.
3. Add 30 µL of the Profiler Plus or COfiler Master Mix to each of the labeled tubes and add approximately 50
 l of mineral oil to each tube. Cap all tubes. Label tubes appropriately.
4. Add the template DNA making sure that the DNA is deposited below the oil. Target the [DNA] to be 1-2.5 ng and the final volume to be 20

L (see the PCR Amplification Sheet). Centrifuge all tubes briefly.
5. Place the tubes in the pre-warmed Thermal Cycler, seating each tube tightly. Set program to run
File # 86.
Cycle Description Temperature Time
Initial Incubation: 95 0 C
Denature 94
0
C
Anneal
Extend
59 0 C
72
0
C
Final Extension
Final Soak
60 0 C
15
0
C
11 Minutes
1 Minute
1 Minute
1 Minute
120 minutes
Indefinitely
Number of Cycles
1
28
1
T/C Serial No. _________________
6. After the first cycle check to see that the samples are seated tightly in the wells.
7. After amplification, remove the tubes from the thermal cycler. Place the products in 4
0
C storage.
Proceed to CE analysis. For long-term storage place the products in the freezer.