Mapping 250K/500K SNP assay

88 GeneChip ® Mapping 500K Assay Manual
PREPARE DILUTED ALIQUOTS OF PURIFIED SAMPLE
Two of the wells on each optical plate must be set up as blanks
containing AccuGENE® water only.
The 12-channel P20 pipette must be accurate to within ± 5%.
To prepare three diluted aliquots of the purified samples:
1. Pour 75 mL of room temperature AccuGENE® water into the
solution basin.
2. Using a 12-channel P200 pipette, aliquot 198 µL of water to:
A. Each well of optical plates 1, 2 and 3.
B. The first four rows of optical plate 4.
3. Using a 12-channel P20 pipette:
A. Transfer 2 µL of each purified PCR product from rows A
through G of the purified sample plate to the corresponding
rows and wells of optical plates 1, 2 and 3 (see Figure 4.5 on
page 89).
B. Pipette up and down 2 times after each transfer to ensure that
all of the product is dispensed.
C. Examine the pipette tips and aliquots before and after each
dispense to ensure that exactly 2 µL has been transferred.
D. Transfer 2 µL of each purified PCR product from row H of the
purified sample plate to the corresponding rows and wells of
optical plate 4.
E. Again, pipette up and down 2 times after each transfer, and
examine the pipette tips and aliquots before and after each
dispense.
The result is a 100-fold dilution.
Two of the wells containing water only will serve as blanks.
4. Set a 12-channel P200 pipette to 180 µL.
5. Mix each sample by pipetting up and down 5 to 10 times.
Be careful not to scratch the bottom of the plate, or to introduce
air bubbles.