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AAT Bioquest

FAM Dye qPCR Calibration Solution *10,000X*

ABI 7500 FAST system FAM dye spectrum.
ABI 7500 FAST system FAM dye spectrum.
ABI 7500 FAST system FAM dye spectrum.
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Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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Physical properties
Molecular weightN/A
SolventWater
Spectral properties
Correction Factor (260 nm)0.32
Correction Factor (280 nm)0.178
Extinction coefficient (cm -1 M -1)83000
Excitation (nm)493
Emission (nm)517
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


Molecular weight
N/A
Correction Factor (260 nm)
0.32
Correction Factor (280 nm)
0.178
Extinction coefficient (cm -1 M -1)
83000
Excitation (nm)
493
Emission (nm)
517
Real-time thermal cyclers require calibration to generate a spectral profile for the fluorescence signal. Since instruments can differ in fluorescence detection sensitivity, the concentrations needed to calibrate any dye, such as FAM, will depend on the device used. The AAT Bioquest FAM dye qPCR Calibration Solution supplied as a 10,000X concentrate is compatible with a wide variety of qPCR instruments, including the ABI7500 Fast, QuantStudio™, and ViiA™ 7 systems. Calibration and verification should be run every six months or as needed. Please refer to the instrument maintenance guide for complete instructions on use.

Platform


qPCR

Recommended platePCR Microplate
Instrument specification(s)FAM Filter

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

This generic protocol is suitable for the ABI7500 FAST qPCR instrument. For complete instructions on use, refer to the maintenance guide for your instrument.

Prepare a FAM calibration plate:
  1. Prepare a FAM Dye qPCR Calibration working solution by mixing 1 µL of the 10,000X FAM Dye qPCR Calibration Solution with 10 mL of nuclease-free water or qPCR buffer. Note the final dye concentration must be optimized based on the calibration requirements for your instrument. Refer to the manufacturer’s maintenance guide for detailed instructions.

  2. Add 20 µL of the FAM Dye qPCR Calibration working solution to all wells of a reaction plate. Note the volume may vary depending on your instrument. Refer to the manufacturer’s maintenance guide for detailed instructions.

  3. Seal the plate with an optical adhesive cover.
  4. Centrifuge the plate for 2 minutes at < 1500 rpm.
  5. Check that the liquid in each well of the plate is at the bottom of the well. If not, centrifuge the plate again at a slightly higher rpm and for a longer period of time.
  6. For complete calibration instructions, refer to the maintenance guide for your instrument.

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.32
Correction Factor (280 nm)0.178
Extinction coefficient (cm -1 M -1)83000
Excitation (nm)493
Emission (nm)517

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