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Genotyping by PCR of a Large Colony of Mice

PCR Success Story #13

Project Description

Mouse Genotyping by PCR of a large mice colony

Mouse Genotyping by PCR works well in their lab, but costs 1000$ for 1000 units of Titanium Taq 🙁

This project illustrates one of the main reasons why we offer FREE PCR services:

Researchers don’t like to test Taq samples for many reasons:

  • too many suppliers;
  • they all say their Taq will work;
  • it requires time to perform tests and performance comparison.

This well-known researcher who’s lab is located at McGill’s Goodman Research Center in Montreal had been using a really expensive Taq DNA Polymerase for many years. But now is the time for change. We offered them our free PCR service. They provided us with protocols, DNA primers and mouse tail gDNA samples (not purified). Please find bellow what we have accomplished using Transgen Biotech’s TransStart TopTaq DNA Polymerase. Because this Mouse Genotyping by PCR project was large, we will not provide PCR setup and cycling protocols for all genes and for simplicity reasons, we will not provide all primer sequences.

The cost for TransStart TopTaq DNA Polymerase currently is 894$ for 3000 units, thereby reducing the researchers cost for genotyping his mice colony by more than 66% without affecting the reliability of their PCR experiments.

 

 

Project Details

Client: McGill University
Date: May 5th, 2016
Type of experiment: Mouse Genotyping by PCR on gDNA
DNA Polymerase: TransStart TopTaq DNA Polymerase

Competitor: Clontech Titanium Taq (not shown)

Gene 1 and 2 Mouse Genotyping with TopTaq

Gene 1 and Gene 2 Genotyping by PCR with TopTaq

PCR amplification of Gene 1 and Gene 2 from transgenic mice gDNA samples.

We followed the lab’s protocol using TransStart TopTaq. However, likely due to the ”quick ‘n dirty” extraction method, some variability was observed in the DNA content of some samples.

floxed Gene 1and Gene 2 Genotyping by PCR

PCR amplification of floxed Gene 1 and Gene 2 alleles from transgenic mice gDNA samples.

We followed the lab’s protocol using TransStart TopTaq. However, likely due to the ”quick ‘n dirty” extraction method, some variability was observed in the DNA content of some samples. The presence of the Cre allele was also assesed, however this reaction was setup separately and too much Taq was used. See Cre in other PCR success stories – no problem at all 🙂

Gene1flox-Gene2flox-Cre Mouse Genotyping with TopTaq
Gene 3, 4 and 5 Mouse Genotyping with TopTaq

Gene 3, 4 and 5 Genotyping by PCR with TopTaq

PCR amplification of Gene 3, 4 and 5 from transgenic mice gDNA samples.

We followed the lab’s protocol using TransStart TopTaq. However,the lab’s protocol for Gene 4 contained half normal concentrations of DNA primers and DNA template – we recommended them to adjust these concentrations to ‘normal’ levels (200 nM each) for the future. Alternatively, there may have been variability in gDNA content in these samples due to the quick ‘n dirty DNA extraction method.

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