Pre-analysis consulting. Consulting services are available including protocols, discussion of sample types, sample number, genes of interest and overall goals of the project.
Laser Capture Microdissection. The facility offers a Zeiss UV-A, nitrogen pulse laser microdissection and catapulting system. This enables specific areas or single cell isolations from different types of tissues and cell cultures. Please contact the facility for further information regarding proper tissue preparation techniques.
Sample preparation. Assistance in sample preparation is available including protocols and provision of sample stabilization solution.
Extractions and cDNA synthesis. gDNA and total RNA extractions for research projects are done in a 96-well based format using Qiagen BioSprint. Additionally researchers can request to have their samples precipitated and cDNA synthesized. A housekeeping gene validation experiment is run prior to profiling in order to determine the most stably transcribed housekeeping gene.
Special protocols. Enhanced RNA or gene-specific pre-amplification protocols are available for projects with low DNA or RNA concentration.
PCR system design. If the gene of interest has been sequenced the facility can provide custom primer design through Primer Express 3 for research projects. Inventoried primers can also be purchased directly from Applied Biosystems.
Assays. More than 3,000 Real-time PCR systems are currently available in the facility including but not limited to: Apple, Chicken, Citrus, Dog, Horse, Cat, Human, Mouse, non-human Primate, Rat, Sturgeon, Tobacco and Trout. Assays can also be purchased. Please check our assays page for availability.
PCR. Complete PCR analysis is done using a laser 7900 HT platform. RNA and DNA 384-well bar-coded plates may be submitted from other labs. Plates from external users will NOT be accepted on Fridays or a day before a university holiday.
Final calculations. Results can be reported either by raw Cq values, using absolute standard methods or the comparative Cq method for relative gene transcription.
Custom design assays. The facility offers full disclosure of primer and probe sequences for newly designed assays. The facility can also provide a written Materials & Methods section for publications for preliminary data, microdissection and pre-amplification.
SNP. SNP genotyping analysis identifies single nucleotide polymorphisms, which are responsible for differences in genetic traits, susceptibility to disease or response to drug therapies. Over 4.5 million human and 10 thousand mouse SNP ready to order assays are available from Applied Biosystems.
A post-PCR analysis method that incorporates the use of high saturation dye for genotyping, mutation scanning and sequence matching. Currently available only in 384-well plate format.
HRM review article (PDF)