Mapping 250K/500K SNP assay

130 GeneChip ® Mapping 500K Assay Manual
HYBRIDIZING SAMPLES USING HEAT BLOCKS
About this Procedure
The following instructions require 2 operators working
simultaneously, each processing two samples at a time. Batches of
sixteen samples at a time are denatured and loaded onto arrays.
Two heat blocks are required: one set to 99 °C; the other set to 49 °C.
Load Samples Onto a Heat Block
1. If the heat blocks are not turned on, preheat them now (set one to
99 °C; the other to 49 °C).
2. Add 190 µL of Hybridization Master Mix to each 1.5 mL
Eppendorf Safe-Lock tube.
3. Transfer the labeled sample from the reaction plate to an
Eppendorf tube containing Hybridization Master Mix (one sample
per tube).
The total volume is now 260 µL.
Reagent Volume/Sample
Hybridization Master Mix 190 µL
Labeled DNA 70 µL
Total 260 µL
4. Vortex at high speed 3 times, 1 sec each time.
5. Pulse spin for 3 sec.
6. Do one of the following:
• If denaturing and loading samples onto arrays now, place the
tubes on ice.
• If not proceeding to denature and hybridization at this time,
store the samples at –20 °C (the mix will not freeze).
7. Place the tubes in batches of 16 at a time onto a heat block as
follows: