GENOME TECHNOLOGY ACCESS CENTER • DEPARTMENT OF GENETICS • WASHINGTON UNIVERSITY SCHOOL OF MEDICINE

PCR Services

 

General Information

GTAC employs PCR for targeted sequencing, SNP genotyping, gene expression and “digital droplet” PCR. Our instrumentation allow us to be able to process a wide variety of both numbers and types of samples. These include:

  • Fluidigm Biomark HD usew integrated fluidic circuits (IFCs) to automate qPCR reactions in nanoliter volumes. This means using less sample and reagent in a single microfluidic device. The Biomark HD system runs IFCs in either real-time or end-point read modes for economical PCR solutions to a range of applications.
  • Fluidigm Juno is capable of custom amplicon-library generation, handling up to 192 samples by 24 reaction wells.
  • Biorad QX200 Droplet Digital PCR system provides quantification of target DNA or RNA molecules for EvaGreen or probe-based digital PCR applications.
  • Biorad CFX96 is a six-channel (five colors and one FRET channel) real-time PCR instrument for single-plex or multi-plex reactions.

Types Of PCR Services

Gene Expression:

Using Taqman probes or custom primers and Evagreen, a DNA intercalating dye, the relative expression of your gene or genes of interest can be quantitatively measured using our services.  High through-put projects are more manageable with the BioMark™ HD System.  This technology allows our group to process samples and assays faster and cheaper than standard plate-based PCR by processing either 48, 96, or 192 samples against 24, 48, or 96 Taqman assays for either 2304, 4608, or 9216 assays per chip.

Single-Cell Gene Expression:

Differential expression levels within individual cells can be quantitatively measured using the BioMark™ HD System.  Cells are processed either in 48 or 96 sample batches for rapid through-put of samples.  The chips can multiplex 48 to 96 assays per sample.  The samples can be evaluated with either Taqman probes or Evagreen intercalating dye.

Copy Number Variation:

The differences in genetic variation and possible association with various diseases can be detected with two types of platforms. Depending upon the sensitivity your project requires, digital PCR is available using the BioMark™ HD System or BioRad QX200 system.

SNP Genotyping:

Genetic variations of single nucleotide polymorphisms can be detected using Taqman probes.  High through-put projects are more manageable with the BioMark™ HD System. This technology allows our group to process samples and assays faster and cheaper than standard plate-based qPCR by processing up to 24, 48, or 96 assays per sample.

Custom Amplicon Sequencing:

Amplicon sequencing consists of sequencing known regions of interest.  This highly targeted approach enables a wide range of applications for discovering, validating, and screening genetic variants in a rapid and efficient manner. This is accomplished by using the multiplexing capabilities of the Fluidigm Juno and Access Array systems.