PACE® Genotyping Assay

About

PACE (PCR Allelic Competitive Extension) genotyping chemistry is a homogeneous, PCR-based allele-specific technology for the analysis of DNA sequence variants, most commonly SNPs (Single Nucleotide Polymorphisms) and Indels (insertion / deletions).

PACE genotyping chemistry is comprised of two parts:

1. PACE Genotyping Assay: two allele-specific forward primers and one common, reverse primer. The allele-specific forward primers each have different 3′ terminal bases reflecting the target variant, and a unique 5’ tail sequence which is incorporated as part of the fluorescent signal mechanism.
2. PACE Genotyping Master Mix: containing all remaining components required for PCR and the generation of fluorescent signals. PACE Genotyping Master Mix contains a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals (FAM and HEX) corresponding to the assay genotypes.

When combined with sample DNA, these components create a PACE Genotyping Reaction, as illustrated in the figure below.

We have extensive knowledge and experience in assay design, especially when it comes to allele-specific PCR. PACE Genotyping Assays are available to purchase either Validated and Unvalidated. Validated assays require customer DNA to validate and optimise, for guaranteed performance. Unvalidated assays are designed in silico and supplied untested.

REQUIRED COMPONENTS

• qPCR machine or Thermocycler + Fluorescent plate reader
• PCR plate or equivalent and appropriate optically clear seal
• Template DNA
• PCR-grade water
• PACE Genotyping Master Mix or PACE 2.0 Genotyping Master Mix

STEPS TO YOUR PACE GENOTYPING ASSAY DESIGN

1. Place your order on this page. Our support team will contact you by email.
2. Fill out an Assay Design Template form with all the information we need to process your custom PACE Genotyping Assay order. We will email you a copy of the template when we first contact you, or your can download a copy here.
3. Email your completed Assay Design Template to support@3crbio.com
4. Using the information you provide us, we will create your PACE Genotyping Assay designs, order the oligos, and send you design sequences.
5. Once we receive the oligos, we assemble the assay(s) and then ship an aliquot to you (unvalidated) or test on your DNA samples before shipping the aliquot to you (validated).