Mapping 250K/500K SNP assay

chapter 4 | 96-Well Plate Protocol 111
3. Pulse spin the enzyme for 3 sec; then immediately place back in
the cooler.
4. Add the TdT enzyme to the master mix.
5. Vortex the master mix at high speed 3 times, 1 sec each time.
6. Pulse spin for 3 sec.
7. Immediately proceed to the next set of steps, Add the Labeling
Master Mix to the Samples.
Table 4.33
Labeling Master Mix
Reagent 1 Sample 96 Samples
(15% extra)
TdT Buffer (5X) 14 µL 1545.6 µL
GeneChip ® DNA Labeling
Reagent (30 mM)
2 µL 220.8 µL
TdT enzyme (30 U/µL) 3.5 µL 386.4 µL
Total 19.5 µL 2152.8 µL
ADD THE LABELING MASTER MIX TO THE SAMPLES
To add the Labeling Master Mix to the samples:
Keep samples in the cooling chamber and all tubes on ice when
making additions.
1. Aliquot 178 µL of Labeling Master Mix to each tube of the strip
tubes.
2. Add the Labeling Master Mix as follows:
A. Using a 12-channel P20 pipette, aliquot 19.5 µL of Labeling
Master Mix to each sample.
B. Pipette up and down one time to ensure that all of the mix is
added to the samples.
The total volume in each well is now 70 µL.