PCR Core Facility

The quantitative PCR core facility in the division of Endocrinology at UCDenver has been in operation since 1998 as a service facility to more than 100 investigators on and off campus. The core facility uses the QuantStudio 7 Flex Real Time PCR instrument from AppliedBiosystems. Acknowledged as the gold standard in q-PCR, the QuantStudio 7 Flex combined with the TaqMan genomic assays provides high throughput and flexibility, allowing users to pursue projects beyond the scope of any other Real time instrument. In collaboration with the Animal facility on campus, highly experienced core personnel have designed and optimized assays to detect sensitive levels of Corynebacterium bovis.

The dedicated core lab space utilizes sterile equipment and aseptic techniques to provide quality data with a level of certainty that is useful for diagnostic research. Furthermore, an efficient turnaround time of 3-4 business days avoids potential delays in data progression.

  • RNA isolation from tissues, cultured cells, blood, fine needle aspiration biopsies and LCM
  • Primer design and optimization 
  • Quantitation of gene expression at the mRNA level
  • DNA copy number assay
  • Micro RNA quantitation
  • SNP genotyping
  • Pathway-specific gene expression profiling
  • ChIP assay end points
  • Plate run (Use of PCR machine)
  • Measurement of absolute telomere length
  • qPCR genotyping assay for Down Syndrome mouse model Ts65Dn
  • C. Bovis detection and quantitation from buccal swab (mouse)
  • MPV/MVM detection and quantitation from fecal pellets (mouse)
  • Mouse Noro Virus (MNV) detection and quantitation from fecal pellets

You may schedule a time to meet at the core facility to discuss the project or

Take the following steps:

  1. Send the mRNA sequence, GenBank acc number or the name of the target specifying the species to vibha.sharma@cuanschutz.edu 
  2. We will design the primers/probe send you the optimal set along with the amplicon seq. You can then blast the amplicon sequence for specificity.
  3. We will place the order and contact you when the primers are received (approximately 7-10 days from ABI).
  4. You will need to provide a positive control in order to optimize the primers/probe and to set up a standard curve for your assay. You may either use an absolute std, relative std po absolute PCR std:
    1. Quantitated riboprobe (absolute standard) or invitro transcribed RNA
    2. RNA from source with known expression of gene (relative standard)
    3. Plasmid-DNA (absolute PCR standard). If you are not sure about a positive control, then we can place the order for the amplicon as a single stranded DNA and use it for std curve.
  5. When your test samples and stds (RNA in water ~ 1 ug/ul) are ready, contact the core via email to set up a date and time for sample drop off.  You can get guidance from the core for nucleic acid isolation, purification and checking the integrity of RNA or DNA.
  6. The results will be emailed to you within 5 business days.
The results of your experiment will include the following: Plots showing the amplification of target and normalizing genes, the standard curve and quantities of target genes normalized to control genes.
1

Send in the mRNA sequence, GenBank acc number or the name of the target specifying the species.

Please email the information to:
vibha.sharma@cuanschutz.edu
2

We will design the primers/probe send you the optimal set along with the amplicon seq.

You can then blast the amplicon sequence for specificity.
3

We will place the order and contact you when the primers are received.

Approximately 7-10 days from ABI.
4

You will need to provide a positive control in order to optimize the primers/probe and to set up a standard curve for your assay.

You may either use an absolute std, relative std po absolute PCR std:
  1. Quantitated riboprobe (absolute standard) or invitro transcribed RNA 
  2. RNA from source with known expression of gene (relative standard) 
  3. Plasmid-DNA (absolute PCR standard). If you are not sure about a positive control, then we can place the order for the amplicon as a single stranded DNA and use it for std curve.
5

When your test samples and stds (RNA in water ~ 1 ug/ul) are ready, contact us at 303-724-3965 to set up a date and time for sample drop off.

You can get guidance from the core for nucleic acid isolation, purification and checking the integrity of RNA or DNA.
Call 303-724-3965
6

The results of your experiment will include the following:

Plots showing the amplification of target and normalizing genes, the standard curve and quantities of target genes normalized to control genes.
7

The results will be emailed to you within 5 business days.

Effective 07/01/2022

SamplePrice
RNA Duplicate$ 30
DNA Duplicate$ 20
rRNA Control $ 35
Standard Curve$ 312
Optimization$ 261
Plate Run$ 172
RNA Extraction-5 samples$ 223
RNA Extraction-10 samples$ 334
RNA Extraction-15 samples$ 416
RNA Extraction-20 samples$ 553
Rodent Pathogen PCR Assay- C. Bovis$ 69
Rodent Pathogen PCR Assay- PARVO$ 79

 

Director:
Bryan Haugen, MD

Bryan.Haugen@cuanschutz.edu

Manager:
Vibha Sharma, PhD

vibha.sharma@cuanschutz.edu 

PRA:
Jacob Calhoun

jacob.calhoun@cuanschutz.edu 

Location:
RCI South, Room 7212
Phone: 303-724-3965