Mapping 250K/500K SNP assay

264 GeneChip ® Mapping 500K Assay Manual
cover, vortex at medium speed for 2 seconds, and spin at
2,000 rpm for 1 minute.
4. Consolidate three PCR reactions for each sample into one well of
the Clean-Up Plate.
For PCR samples prepared in three 96-well PCR plates, an 8- or 12channel
pipette can be used to transfer each row of 12 samples in
the PCR plates to the corresponding row of the Clean-Up Plate.
With the vacuum on, the three PCR reactions for each sample
(300 µL) can be combined into one well of the Clean-Up Plate.
Be sure to dilute EDTA to 0.1 M. A higher concentration may
interfere with downstream steps.
To avoid piercing the membrane, do not pipet up and down in the
Clean-Up Plate.
Make sure the orientations of PCR plates are consistent.
Use a method of distinguishing between used and unused wells on
the plate.
Be sure to maintain the vacuum at 600 mbar.
Three water washes must be done to remove PCR reaction
contaminants (e.g., EDTA). Be sure to completely dry the
membrane between each wash.
After the final wash, every well must be completely dry before
adding elution buffer. This step is critical to prevent the dilution of
DNA with water.
5. Apply a vacuum and maintain at ~600 mbar until the wells are
completely dry.