Mapping 250K/500K SNP assay

58 GeneChip ® Mapping 500K Assay Manual
Table 4.12
Sty I Ligation Master Mix
Reagent 1 Sample 96 Samples
(15% extra)
Adaptor Sty I (50 µM) 0.75 µL 82.8 µL
T4 Ligase Buffer (10X) 2.5 µL 276 µL
T4 DNA Ligase (400U/µL) 2 µL 220.8 µL
Total 5.25 µL 579.6 µL
ADD LIGATION MASTER MIX TO REACTIONS
To add Ligation Master Mix to samples:
1. Using a single channel P100 pipette, aliquot 48 µL of Ligation
Master Mix to each tube of the strip tubes on ice.
2. Using a 12-channel P20 pipette, aliquot 5.25 µL of Ligation
Master Mix to each reaction on the Digestion Stage Plate.
Reagent Volume/Sample
Digested DNA 19.75 µL
Ligation Master Mix* 5.25 µL
Total 25 µL
* Contains ATP and DTT. Keep on ice.
3. Seal the plate tightly with adhesive film.
4. Vortex the center of the plate at high speed for 3 sec.
5. Spin down the plate at 2000 rpm for 30 sec.
6. Ensure that the thermal cycler lid is preheated.
7. Load the plate onto the thermal cycler and run the 500K Ligate
program.