Services - Real Time PCR

The facility has a iCycler iQ Real-Time Detection System (BioRad) which can accurately quantitate PCR products in real time. The iQ system is very sensitive and can simultaneously measure up to 4 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 iQ system can analyze all of the current detection chemistries, such as 5xBR Green, Tagman, Molecular Beacons, etc.

The service includes the use of the iCycler iQ system and a file with the results. In addition the Facility can provide a copy of the iQ 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.

The iCycler iQ Real-Time Detection System can be reserved through dnaLIMS, so click on the link below to (1) create an account if you have not done so previously, and (2) then click on coreCal to view when the instrument is available.

dnaLIMS

The following quantitative real-time PCR reagents are available at the facility:

  1. BioRad iQ SYBR Green Supermix (#170-8880 )
Customer files
(Copy your files from this secure server site that requires an assigned user name and password.)

Many of the links below are pdf files and they require Acrobat Reader to be viewed. Acrobat Reader can
be obtained free of charge from Adobe, Inc

 

References for research done at PMGF:

  1. Hanson T. and F. R. Tabita. 2003. Photo. Res.
  2. Sylvester, J. et al. 2004. J. Nutr.

BioRad Bulletins:

  1. iCycler iQ Real-Time Detection System manual [pdf file]
  2. Real-time PCR: General Considerations [pdf file]
  3. Guide to designing primers and probes: Taqman and Molecular Beacon [pdf file]
  4. Real-time PCR using the iCycler iQ Detection system and Intercalation Dyes [pdf file]
  5. The iCycler iQ Detection System for Taqman Assays [pdf file]
  6. The iCycler iQ Detection System for Multiple Real-Time PCR Assays [pdf file]

Qiagen Bulletins:

  1. Qiagen SYBR Green Manual [pdf file]

Applied Biosystems Bulletins:

  1. Guide to designing Taqman probe [pdf file]
  2. Consumables for real-time PCR (including figures that demonstrate how Taqman probes and SYBR Green work) [pdf file]