Real-time Quantitative PCR

diagram of pcr amplificationThe 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:

  1. Go to dnaLIMS and login
    (or create an account if you have not done so previously)
  2. Click on the Tab "Other Services"
  3. Click on "Real-time PCR" and then follow the instructions
  4. Contact us at pmgf@osu.edu or 614-247-6204 to schedule an appointment

How to deliver samples

Instructions to deliver samples from both campus and non-campus clients.

How to get results:

  1. results will be sent by email immediately after the run is complete
  2. clients will receive a .pcrd file (proprietary with all data) and summary of the run with results in pdf format

References and Resources


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.


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