2.3. APOE genotyping

        Materials Needed:

        Sample to SNP Kit (ThermoFisher; Applied Biosystems-4403087)

        TaqMan GTXpress™ Master Mix (ThermoFisher 4401892)

        Taqman probe rs429358 (C___3084793_20) 

        Taqman probe rs7412 (C____904973_10)

1. Use sample to SNP kit to isolate DNA. Incubate 2 μL of whole blood with 20 μL of lysis buffer for 3 minutes at room temperature. Add 20 μL DNA stabilizer solution. Sample is ready for allelic discrimination assay. Samples can be processed using microfuge tubes or 96 well plate (depending on number). Ensure tubes are labeled for storage. 
2. Set up allelic discrimination assay using 5 μL TaqMan GTXpress™ Master Mix, 2.5 μL water, 2μL DNA sample, and 0.5 μL probe. 
3. For APOE genotyping we use SNP IDs:  rs429358 (C___3084793_20) and rs7412 (C____904973_10). 
4. Use the following PCR parameters in the qPCR machine (Step One Plus): 

        95 °C for 20 seconds, 40 cycles of [95 °C for 3 seconds and 60 °C for 20 seconds] 

    5. Use positive and negative control samples.