QGCL Assays and Methods

Assay List
The QGCL has a large number of real-time qPCR assays available for mostly mouse, rat and human species. The current assay list can be downloaded as a PDF. Should you not find the transcript or gene you are looking for on the list, contact Dr. Shipley about having a new real-time qPCR assay developed.

DNase I treatment of Total RNA
Total RNA samples have to be treated with RNase-free DNase I. The real-time assays are not purposely designed across Exon junctions. In the early days, this information was not always available. Most importantly, most transcripts have pseudogenes, unexpressed version of transcripts without junctions, in the DNA which will amplify in any case.
(DNase I protocol)

RNA Isolation for qPCR
Preparation of total RNA from tissue culture cells for qPCR analysis is best done over a column-based kit such as Qiagen’s RNEasy, Clonetech’s Nucleobond or the Ambion RNAqueous kit. From whole tissue, it is best to start with a phenol-based guanidinium buffer such as Tri-Reagent, RNAsol, or one of the many other similar reagents followed by RNA isolation over a Qiagen column (RNA-TriReagent_Qiagen). For fixed and embedded material, we have had good luck with the Epicentre Masterpure RNA isolation kit.

Sample Submission Form
Samples must be accompanied by a sample submission form. Please fill it out completely and bring it with you when you drop off samples for analysis.

Contact Dr. Shipley for current Core Lab charges.