FAM/HEX Dual Channel Calibrator Set

Highlights

• Calibrates the Open qPCR dual channel instrument
• Lyophilized format for ambient temperature shipping
• Provides 3 calibrations

Description

The FAM + HEX Calibrator Set contains everything necessary to calibrate the Open qPCR dual channel instrument. The calibration process compensates for well-to-well variations to ensure accuracy and reliability in the amplification results.

Chai's FAM and HEX calibrators are specially designed 10-mer DNA oligos with the fluorophore covalently linked to the 5’ end. Because certain reaction chemistries result in end products where the fluorophore remains attached to the probe (i.e. Scorpion probes), and other chemistries which cleave the fluorophore (i.e. TaqMan) generally leave a few bases attached to it, the 10-mer calibrators closely reflect actual reaction conditions.

Normalized spectrum profiles of: a. HEX Calibrator in Chai's 1X Reconstitution Buffer. b. FAM Calibrator in Chai's 1X Reconstitution Buffer. c. Amplicons generated from end-point PCR using HEX probe in Chai's PCR Master Mix 2X. d. Amplicons generated from end-point PCR using FAM probe in Chai's PCR Master Mix 2X. All four reactions were read at 60 ºC on a spectrophotometer.

Specifications

Reconstitution Procedures

1. Prior to opening the calibrator tubes, use a mini centrifuge to spin down the two lyophilized FAM and HEX vials. It is important to gather all materials at the bottom of the tube.
2. Using a sterile tip, pipette 1.32 mL of the reconstitution buffer into each of the calibrator vials. Mix well by pipetting the solution up and down ten times before capping the vials. Alternatively, ensure vial caps are tightly secured before vortexing the vials for 5-10 seconds to ensure homogenous mixing.
3. Incubate the vials at room temperature for at least thirty minutes. This is necessary to ensure that the product has completely dissolved. During the thirty-minute incubation period, protect the calibrator vials from light by placing them in an opaque drawer or covering them with aluminum foil.
4. Following the thirty-minute incubation, open the vial caps and use a sterile tip to pipette up and down ten times. Alternatively, ensure vial caps are tightly secured before vortexing the vials for 5-10 seconds to ensure homogenous mixing.
5. Spin down the vials using a mini centrifuge to collect all liquid contents at the bottom of the tube. If a centrifuge is unavailable, flick the tubes in a downward motion to gather liquid contents at the bottom. The reconstituted calibration vials are now ready for use.

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