Mapping 250K/500K SNP assay

280 GeneChip ® Mapping 500K Assay Manual
2. Mix well.
This Hybridization Cocktail Master Mix can be stored at –20°C
before proceeding to the next step.
3. Transfer each of the labeled samples from the plate to a 1.5 mL
Eppendorf tube. Aliquot 190 µL of the Hybridization Cocktail
Master Mix into the 70 µL of labeled DNA samples as follows:
Table C.15
Reagent Volume/Rx
Labeled DNA 70 µL
Hybridization Mix 190 µL
Total 260 µL
4. Heat the 260 µL of hybridization mix and labeled DNA at 99°C
in a heat block for exactly 10 minutes to denature.
Denaturation of the labeled DNA sample is important to maximize
binding to the oligonucleotides on the array surface. The sample
must be heated to at least 95°C for 10 minutes prior to adding the
DNA to the array. If a precipitate forms during this step, resuspend
it before hybridization. Hybridize the arrays for at least 16 hours at
49°C. This temperature has been optimized for this product, and
should be stringently followed.
5. Cool on crushed ice for 10 seconds.
To avoid the formation of aggregates, do not leave on ice for longer
than 10 seconds.
6. Spin briefly at 2,000 rpm in a microfuge to collect any condensate.
7. Place the tubes at 49°C for 1 minute.