Mapping 250K/500K SNP assay

appendix C | Low Throughput Protocol 271
Table C.10
Diluting the Fragmentation Reagent – Combine Reagents in the Order Shown
Reagent 2 units/µL 3 units/µL
H 2O, Molecular Biology Grade 105 µL 106 µL
10X Fragmentation Buffer 12 µL 12 µL
Fragmentation Reagent* 3 µL 2 µL
Total 120 µL 120 µL
* Add the Fragmentation Reagent last, after allowing the water and buffer to
cool on ice.
If the concentration on your tube is not shown in the table above,
use the formula provided in Step 3.
5. Quickly and ON ICE, divide the diluted Fragmentation Reagent
into 8 or 12 microtube strips.
6. Add 5 µL of diluted Fragmentation Reagent (0.05 U/µL) with an
8- or 12-channel pipette to the fragmentation plate containing
Fragmentation Mix ON ICE. Pipet up and down several times to
mix. Be sure to change tips between samples. The total volume for
each sample is listed below.
Table C.11
Reagent Volume/Sample
Fragmentation Mix 50 µL
Diluted Fragmentation Reagent (0.05 U/µL) 5 µL
Total 55 µL
For 90 µg of purified PCR product, a total of 0.25 U of Fragmentation
Reagent is needed in a final reaction volume of 55 µL.
7. Cover the fragmentation plate with a plate cover and seal tightly.