Mapping 250K/500K SNP assay

124 GeneChip ® Mapping 500K Assay Manual
HYBRIDIZING SAMPLES USING A THERMAL CYCLER
Method 1 — Using a GeneAmp® PCR System 9700
The thermal cycler used for this method must be a GeneAmp PCR
System 9700 located adjacent to the hybridization ovens. This
particular thermal cycler is required because of the way the lid
operates. You can slide it back one row at a time as samples are loaded
onto arrays. Keeping the remaining rows covered prevents
condensation in the wells.
Add Hybridization Master Mix and Denature the Samples
To add Hybridization Master Mix and denature the samples:
1. Pour 20.9 mL Hybridization Master Mix into a solution basin.
2. Using a 12-channel P200 pipette, add 190 µL of Hybridization
Master Mix to each sample on the Label Plate.
Total volume in each well is 260 µL.
3. IMPORTANT Seal the plate tightly with adhesive film.
4. Vortex the center of the plate for 3 minutes.
5. Spin down the plate at 2000 rpm for 30 sec.
6. Cut the adhesive film between each row of samples.
Do not remove the film.
7. Place the plate onto the thermal cycler and close the lid.
8. Run the 500K Hyb program.
500K Hyb Program
Temperature Time
95ºC 10 minutes
49ºC Hold