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.
HRM requires optimal primers design
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 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.