Real-Time PCR Sample Preparation
Real-Time PCR Fees
Real-Time PCR Resources
Real-time PCR
The IIHG Genomics Division maintains the following models of real-time or PCR instruments:
Applied Biosystems QuantStudio 7 Flex
Applied Biosystem QuantStudio 7 Pro
The instruments have interchangeable blocks that permit the running of 96- and 384-well plate formats. The ABI Model QuantStudio Flex 7 also has an interchangeable block that permits the running of the TaqMan Low Density Array (TLDA) cards.
The Design and Analysis Software v2.3 can be used to analyze data collected from both the QuantStudio 7 Flex and QuantStudio 7 pro instruments.
Real-Time PCR Sample Preparation
- Make a reservation using the online calendaring system.
- Print and complete the Real-Time PCR Request Form.
- Prepare your samples -
- Users are responsible for acquiring the reaction reagents, primers, probe, and reaction vessel. Real-Time PCR reagents and consumables are available from the Biomedical Research Store (2nd Floor EMRB).
- Samples should be prepared in tubes or plates that are ready to load onto the instrument.
- Bring your prepared samples and request form to the IIHG Genomics lab prior to your reservation time.
- We will be responsible for setting up the instrument and data collection.
- Results will be provided via email or can be transferred to a portable drive provided by the investigator.
Real-Time PCR cost per run: $25/plate
Real-Time PCR Resources
Below are links to information which users may want to consider when designing and publishing a real-time or digital PCR experiment.
Real-Time PCR Resources
- The MIQE Guidelines: Minimum Information for Publication of Quantitative PCR Experiments
Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009 Apr;55(4):611-22. doi: 10.1373/clinchem.2008.112797. Epub 2009 Feb 26. PMID: 19246619. 
ABI Prism 7700 Sequence Detection System- Statistical Estimations of PCR Amplification Rates
- Sequence Detection Systems Quantitative Assay Design and Optimization
- DNA/RNA Real-Time Quantitative PCR
- Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems
Bustin SA. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol. 2002 Aug;29(1):23-39. doi: 10.1677/jme.0.0290023. PMID: 12200227. - Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays
Bustin SA. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000 Oct;25(2):169-93. doi: 10.1677/jme.0.0250169. PMID: 11013345.