Mapping 250K/500K SNP assay

chapter 4 | 96-Well Plate Protocol 131
A. Place four tubes onto a heat block at 99 °C and set a timer for
10 minutes.
B. Wait 3 to 4 minutes, then place another 4 tubes onto the heat
block and set another timer for 10 minutes.
C. Repeat this procedure until there are 16 samples loaded onto
the heat block.
Remove Samples from Heat Block and Load Onto Arrays
Two operators will perform this procedure at the same time, two
samples per person.
To load samples onto arrays, 16 samples at a time:
1. When the first timer indicates 10 minutes has transpired:
A. Immediately remove the first samples (two per operator).
B. Cool on crushed ice for 10 sec, then remove immediately.
Cool for 10 sec only. If left on ice longer, aggregates may form.
These aggregates will not break apart at 49 °C and will reduce your
call rate.
Cooling on ice is required for this method only due to the loose fit
of the tubes on the heat blocks. This step helps to ensure that the
samples cool quickly to 49 °C.
2. Pulse spin for 3 sec.
3. Place the tubes back on the heat block at 49 °C for 1 minute.
4. Remove tubes from the heat block, and check for precipitate.
5. Using a single-channel P200 pipette, remove 200 µL of denatured
sample from one tube.
6. Immediately inject the sample into an array.
7. Cover each septa with a Tough-Spot (Figure 4.8 on page 127).
8. Repeat steps 5 through 7 for the next sample.
9. Immediately load the arrays into a hybridization oven tray, 4 arrays
per tray evenly spaced.