Mapping 250K/500K SNP assay

chapter 4 | 96-Well Plate Protocol 129
9. Cover the wells with a fresh strip of adhesive film and place in the
cooling chamber on ice.
10. Remove the next strip of 24 wells and place it on the thermal
cycler.
11. Run the 500K Hyb program.
12. Repeat steps 1 through 11 until all of the samples have been loaded
onto arrays.
Operator 2 Tasks
1. Cover the septa on each array with a Tough-Spot (Figure 4.8).
2. When 4 arrays are loaded and the septa are covered:
A. Load the arrays into an oven tray evenly spaced.
B. Immediately place the tray into the hybridization oven.
Do not allow loaded arrays to sit at room temperature for more
than approximately 1 minute. Ensure that the oven is balanced
as the trays are loaded, and ensure that the trays are rotating at
60 rpm at all times.
Because you are loading 4 arrays per tray, each hybridization
oven will have a total of 32 arrays.
Operators 1 and 2
• Load no more than 32 arrays in one hybridization oven at a time.
• All 96 samples should be loaded within 1 hour.
• Store the remaining samples and any samples not yet hybridized in
a tightly sealed plate at -20 °C.
• Allow the arrays to rotate at 49 °C, 60 rpm for 16 to 18 hours.