The MBCF houses a Roche LC480II Real-Time PCR instrument and can perform a variety qPCR based studies, experiments and assays. We can help guide you in the design of your experiment, sample acquisition, template quality assessment, assay design, plate setup and subsequent data analysis. We also can perform many or all of the steps of your study. Please inquire contact BDCBioResourcesCore by email and we can discuss your project and how we can be of assistance.
In conjunction with the qPCR products and services that the MBCF offers, we also provide RNA quality assessment utilizing the RNA Nano chip on the Bioanalyzer 2100 platform. The quality of RNA template for qPCR applications is critical for the success of your experiment.
Quantitative Real-Time PCR has become a very common technology being utilized for a vast array of research and clinical applications. Rapid advancements in qPCR technologies have necessitated the establishment of scientific consensus and guidelines for qPCR applications. The MIQE Guidelines are a great resource to ensure a solid, standardized experimental approach.
Listed below are a number of helpful links to references and information that will help users for all phases of a quantitative study.
Links for Quantitative Real-Time PCR Resources and Documentation
Clin Chem. 2009 Apr;55(4):611-22. doi: 10.1373/clinchem.2008.112797. Epub 2009 Feb 26.
The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.
Methods. 2010 Apr;50(4):227-30. doi: 10.1016/j.ymeth.2009.11.001. Epub 2009 Dec 5.
How to do successful gene expression analysis using real-time PCR.
Methods. 2010 Apr;50(4):S1-5. doi: 10.1016/j.ymeth.2010.01.005.
A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines.
BMC Mol Biol. 2010 Sep 21;11:74. doi: 10.1186/1471-2199-11-74.
MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments.
Clinical Chemistry. June 2011 vol. 57 no. 6 919-921 Published March 18, 2011. doi: 10.1373/clinchem.2011.162958
Primer Sequence Disclosure: A Clarification of the MIQE Guidelines
Methods of RNA Quality Assessment from Promega
RNA Quality Control and Assessment from BioMedicalGenomics.org
Genome Biology. 2002, 3(7):research0034.1–0034.11 Published: 18 June 2002
Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes
Cancer Res. August 1, 2004 64; 5245 doi: 10.1158/0008-5472.CAN-04-0496
Normalization of Real-Time Quantitative Reverse Transcription-PCR Data: A Model-Based Variance Estimation Approach to Identify Genes Suited for Normalization, Applied to Bladder and Colon Cancer Data Sets
Nucl. Acids Res. (2009) 37 (7):2065-2069.doi: 10.1093/nar/gkp056 First published online: February 17, 2009
RDML: structured language and reporting guidelines for real-time quantitative PCR data
Vender Links for Quantitative Real-Time PCR Resources
BioRad qPCR Bulletin
AB qPCR Overview: Guide to Performing Relative Quantitation of Gene Expression Using Real-Time Quantitative PCR
PCR 4th Edition (2016, 6 Chapters and 68 pages)
Strategene Introduction to Quantitative PCR
High Resolution Melt (HRM) Analysis – A Practical Guide
qPCR Technical Guide Sigma
qPCR guide from IDT
qPCR guide - Eurogentec
Understanding Ct Application Note
In addition to providing molecular services, the Core provides hands-on training of many molecular biology techniques where researchers and their staff can gain basic skills working on projects specifically related to their interests. Consultation services are also provided for advanced training and collaboration. Please contact BDCBioResourcesCore by email or Howard Davidson, PhD (Director) for inquiries regarding your project.
Barbara Davis Center
University of Colorado, Anschutz Medical Campus
1775 Aurora Court, 4th floor, Room 4207
Aurora, CO 80045
Molecular Biology contact personnel:
Mark Farrell, Core Manager