Mapping 250K/500K SNP assay

STEP 5: PCR Purification and Elution with
Clontech Clean-Up Plate
REAGENTS AND EQUIPMENT
appendix C | Low Throughput Protocol 263
• DNA Amplification Clean-Up Kit, to be used with Affymetrix
DNA products: Clontech, P/N 636974 (1 plate) or P/N 636975 (4
plates) (each kit also contains RB Buffer)
• Manifold - QIAvac multiwell unit: QIAGEN, P/N 9014579 1
• EDTA (0.5 M, pH 8.0): Ambion, P/N 9260G
• Molecular Biology Grade Water: Bio Whittaker Molecular
Applications/Cambrex, P/N 51200
•Biomek ® Seal and Sample Aluminum Foil Lids: Beckman, P/N 538619
• Jitterbug 115 VAC: Boekel Scientific, P/N 130000
• Vacuum Regulator for use during the PCR clean up step. QIAGEN
Vacuum Regulator: QIAGEN, P/N 19530*
* The Clontech protocol requires ~600 mb vacuum If your lab does not have an internally
regulated vacuum source, this vacuum regulator is strongly suggested.
Follow the steps as outlined below. Consult the Clontech Clean-Up
Plate Handbook for the general procedure and ordering information.
1. Connect a vacuum manifold to a suitable vacuum source able to
maintain ~600 mbar, e.g., QIAvac Multiwell Unit (QIAGEN).
Place a waste tray inside the base of the manifold.
2. Place a Clean-Up Plate on top of the manifold. Cover wells that are
not needed with PCR plate cover.
To cover the unused wells, a PCR plate cover or an aluminum foil lid
can be placed on top of the Clean-Up Plate. Apply pressure to make
the cover stick to the plate. Cut the adhesive film between the used
and unused wells. Remove the portion that covers the unused wells
you want to use.
3. Add 8 µL 0.1 M EDTA to each PCR reaction. Seal plate with plate
1 ® ® Clontech also supports the Millipore MultiScreen , Promega Vac-Man 96, and Bio-Rad Aurum vacuum manifolds
for use with the Clean-Up Plates.