Mapping 250K/500K SNP assay

PCR STAGING AREA
252 GeneChip ® Mapping 500K Assay Manual
2. Add 5 µL genomic DNA (50 ng/µL) to each well of 96-well plate.
Total amount of genomic DNA is 250 ng for each Restriction
Enzyme.
3. Aliquot 14.75 µL of the Digestion Master Mix to each well of the
96-well plate containing genomic DNA.
Table C.3
Reagent Stock Volume/Sample
Genomic DNA (50 ng/µL) 5 µL
Digestion Master Mix 14.75 µL
Total Volume 19.75 µL
To expedite the aliquoting, the master mix can be first divided into
8 or 12 microwell strips and then dispensed into the plate with an
8-channel or 12-channel pipette. Pipet up and down for several
times to mix the genomic DNA and digestion mix. Be sure to
change tips between samples.
4. Cover the plate with a plate cover and seal tightly, vortex at
medium speed for 2 seconds, and spin at 2,000 rpm for 1 minute.
5. Put the plate on a thermal cycler and run the 500K Digest program:
500K Digest Program
Temperature Time
37ºC 120 minutes
65ºC 20 minutes
4ºC Hold
Store sample at –20°C if not proceeding to the next step.