ABI 7900HT and BioRad CFX384 Real Time PCR Thermocyclers

For access or consultation: Dr. Kim DeWeerd 591-8831 or kdeweerd@albany.edu

The Molecular Core has one ABI 7900HT and one BioRad CFX384 real time PCR thermocyclers.

Purpose:
Real time thermocyclers are used to quantify the amount of specific sequences of DNA in samples. The most common use is to measure the expression levels of specific genes in cells by first converting mRNA to cDNA and then measuring this DNA and comparing it to genes with established expression levels.

Specific Description:
Real time PCR thermocyclers can measure dyes designed to increase or decrease in fluorescence intensity as a product of newly synthesized DNA. Specific genes are targeted using primers that anneal to one or more DNA sequences specific to the gene. The concentration of the DNA sequence associated with a gene is then quantified and compared to the concentration of other targeted genes in the same cells to determine how their ratio changes under different experimental conditions. Both thermocyclers measure fluorescence in 384 well plates. The excitation source for the ABI 7900HT is a 488 nm argon-ion laser. Fluorescence detection occurs with a spectrograph and CCD camera. Multiplexing with several dyes at once is possible in both thermocyclers. The BioRad CFX384 uses five filtered LEDs to excite fluorophores and five filtered photodiodes to detect covering a 450-690 nm emission/excitation range. Each thermocycler has its own specific software to analyze the data.

For More Information:
ABI 7900 HT
CFX-384

Instrument Photographs:
Click to enlarge