The facility has a CFX96 Real-Time Detection System (BioRad) which can accurately quantitate PCR products in real time. The CFX96 system is very sensitive and can simultaneously measure up to 6 different PCR products in each of 96-wells. This system is adept at precisely measuring the concentration of specific RNAs, measuring gene copy number as well as perfoming other applicatons that require the measurement of specific DNA molecules. The CFX96 system can analyze all of the current detection chemistries, such as SYBR Green, Taqman, etc. In addition, the instrument can be used as a specialized fluorometric based plate reader that can detect most dyes for applications such as high resolution melt analysis, single nucleotide polymorphism detection, and measuring protein melting.
The service includes the use of the CFX96 system and a file with the results. In addition, the Facility can provide a copy of the CFX96 software at no cost to you. The Facility will provide the 96-well plate and optical film which is included in the per run cost. The customer will be responsible for providing all other consumables (e.g. primers, polymerase, nucleotides, etc.) and setting up the reactions in the plate.
Order and Instrument Reservation
How to place an order and make a reservation:
- Contact us at firstname.lastname@example.org or 614-247-6204 to place an order and schedule an appointment
How to deliver samples
Instructions to deliver samples from both campus and non-campus clients.
How to get results:
- results will be sent by email immediately after the run is complete
- clients will receive a .pcrd file (proprietary with all data) and summary of the run with results in pdf format
References and Resources
- Procedures and recommendations for Quantitative PCR [pdf]
- Resources for real-time qPCR
- Results for real-time qPCR Instrument and Example
- Hanson T. and F. R. Tabita. 2003. Photo. Res. [pdf]
Insights into the stress response and sulfur metabolism revealed by proteome analysis of a Chlorobium tepidum mutant lacking the Rubisco-like protein
- Sylvester, J. et al. 2004. J. Nutr. [pdf]
Development of an Assay to Quantify Rumen Ciliate Protozoal Biomass in Cows Using Real-Time PCR1
- Lavinder, J. et al. 2009. J. Am. Chem. Soc. (Protein Melting). [pdf]
High-Throughput Thermal Scanning: A General, Rapid Dye-Binding Thermal Shift Screen for Protein Engineering
High Resolution Melt Analysis
The CFX96 can perform genotyping by identifying alleles present, and therefore species as well through High Resolution Melt (HRM) analysis of amplicons. In combination with an intercalating fluorescent dye such as EvaGreen a difference as small as one base can be detected in the shift or difference between the melt curves. With known controls the exact allele, and therfore species or genotype can be identified.
- Order and Instrument Reservation
- Resources for HRM
- Example of results for HRM
- Zianni, M. et al. 2013. J. Biomolec. Tech. [pdf]
[pdf] - Some links on this page are to .pdf files. These are designated by [pdf] following the link. If you require these materials in alternate format contact our web manager.