The Mice Genotyping Service proposed by the department of Veterinary Resources is based on recommendations of Jackson Laboratories and Garvan Institute.
Hold the Agarose! Advanced PCR Methods for Genotyping Mice
Genotyping FAQ - The Jackson Laboratory
Mouse Genotyping | Garvan Institute of Medical Research
Establishing New Mouse Genotyping Projects
To introduce a new genotyping protocol to the service
Follow instructions and
Contact Hagit Dafn
Hagit Dafni
TD-PCR & HRM
PCR is set up in a 384well plate scale with fully automated liquid handling system (Biomek i5; Beckman Coulter)
according to a plan generated by an in-house programming software (developed in MATLAB).
We perform a Touchdown (TD) realtime PCR in combination with High Resolution Meltcurve (HRM) analysis to deduct the genotypes.
Primer Design
HRM requires optimal primers design
Results Analysis
Results analysis is performed using an in-house programming software (developed in MATLAB).
The analysis code is available on demand.
Genotyping reports are posted in a group-dedicated DropBox.
DNA Samples
DNA is extracted from tissue samples using NaOH extraction (quick "dirty" DNA preparation).
Reference: Truett GE et al. 2000. Biotechniques 29(1):52-54.
DNA samples are stored for 2 weeks from posting the results and can be collected for further experiments upon request.