25
o
Mini centrifuge
Reconstitution Procedures
Please note that the lyophilized calibrators may be difficult to see.
1.
Before opening the calibrator tubes, use a mini centrifuge to spin down the lyophilized
HEX and FAM vials for 10 seconds. It is important to gather all materials at the bottom of
the tube.
2.
Using a new tip, pipette 1.32 mL of the provided reconstitution buffer into each of the
calibrator vials. Mix well by pipetting the solution up and down 10 times or by vortexing
the vials for 10 seconds. Make sure to spin down the vials.
3.
Incubate the vials at room temperature for at least 30 minutes. This is necessary to
ensure that the product has completely dissolved. During the 30-minute incubation
period, protect the calibrator vials from light by placing them in an opaque drawer or
covering them with aluminum foil.
4.
After the 30-minute incubation, open the vial caps and use a new tip to pipette up and
down ten times or vortex for 10 seconds.
5.
Spin down the vials using a mini centrifuge to collect all liquid contents at the bottom of
the tube. The reconstituted calibration vials are now ready for use
.
Preparing for Calibration
The FAM and HEX solutions are light sensitive. Keep aliquoted solutions protected from
light. If you are using calibrators that have previously been stored at 4
°
C, you do not
need to equilibrate the solution to room temperature before running the calibration
process. You may use the calibrator solution directly from the fridge.
1.
Obtain 6 new 8-well 100
µ
L PCR tube strips. Check that there are no cracks on the
tubes.
2.
Vortex both the FAM and HEX vials for 10 seconds or invert the vials 10 times to ensure
homogenous mixing of the solution. Spin down the vials.
3.
Aliquot 25
µ
L/tube of the FAM solution into 2 new PCR tube strips. Cap tubes tightly and
spin down.
4.
Aliquot 25
µ
L/tube of the HEX solution into 2 new PCR tube strips. Cap tubes tightly and
spin down.
