Success #2: Mouse Genotyping by PCRPCR Success Story #2
2 bands instead of 1. Wrong band is predominant.
This challenge consisted in optimizing Genotyping by PCR protocol for a well established lab located at the Lady Davis Institute in Montreal. On that day during a visit in their lab, Audrey expressed how she was getting frustrated with a routine mouse DNA genotyping PCR for which she was getting mostly the wrong band size using a well-known competitor Taq DNA polymerase. Cost was an issue, but performance was critical. The researcher provided us with PCR primers and 2 gDNA samples Please find bellow what we have accomplished using Transgen Biotech’s DNA polymerases in a total time of less than 4h (including driving in traffic). What does your Taq supplier do for you?! 🙂
Client: Jewish Hospital – Lady Davis Institute
Date: April 5th, 2016
Type of experiment: Mouse genotyping by PCR
DNA Polymerase: EasyTaq DNA Polymerase.
Competitor: Invitrogen Taq DNA polymerase
Nearly perfect Mouse DNA genotyping on the first attempt
Project #2 took less than 4 hours to solve.
1- Drive to our office in traffic from Montreal
2- Setup and run the PCR reaction,
3- Run the gel. Done.
The customer had given us 2 unpurified mouse gDNA samples and a set or primers. PCR was performed using either EasyTaq, TransTaq HiFi, TransStart TopTaq or TransStart FastPfu. Although the result was satisfying, a little tiny non-specific band ∼480bp was observed. NOTE: this contaminating band was predominant in the customer’s hands using Taq DNA Polymerase from supplier Negortivni. Advice was given to the customer on how she could resolve this issue. See testimonial below.
et je viens de faire mon pcr à 55 et 57, 10 sec annealing –> j’ai une belle bande unique avec ta Taq magique! merci tout plein pour le troubleshooting.