Real Time PCR
Transnetyx uses a quantitative or real time PCR method for genotyping. Using real time not only allows Transnetyx to have a high-throughput, fully automated process but also gives customers a more sensitive and accurate method for measuring and detecting mutations.
Transnetyx will convert your existing gel-based PCR to a real time assay based on the genetic information you provide.
| Click here to view how our
process differs from Traditional PCR
Sending this information to Transnetyx is easy; simply log into your account at www.Transnetyx.com and complete the online submission form. The process takes approximately 7 minutes and provides information about your strain to a Transnetyx Assay Design Specialist so they can design your real time PCR assay.
With more than 10,000 real time PCR assays in stock Transnetyx may already have your assay in house. If this is not the case we'll design a custom assay for your mutation. Either way you will receive email notifications throughout the entire process to give you real time status updates on your assay.
Assay Design Strategy
Transnetyx will design and run one assay per allele associated with the strain in question
- 1 assay for a randomly integrated transgene
- 2 assays for conventional targeted mutations
- 3 (or more) assays for conditional mutations
Assays are targeted based on the type of allele to be detected
- Wild-type alleles will be targeted to the region deleted or disrupted
- Knockout or Knock in alleles will be targeted to the insert or junction of the insert and endogenous mouse DNA
- Point mutations and other small base pair changes will use a multiplex assay targeted to each of the versions of the allele (a SNP type assay)
Once an assay is in stock or assigned to your strain, your samples will be run and the results reported online.
Results are returned as positive or negative for each assay/allele tested. This is similar to a gel-based assay which yields either band or no-band for each allele. A positive (+) result is presence of the allele in question and a negative (-) is absence of that particular allele.
The genotype is interpreted based on the combination of alleles indicated by the above results. Additionally, Transnetyx offers a “Results Translator” that can be set up to interpret the allele based results for the customer.