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Chapter 3 Prepare the digital PCR reaction and load the chips Load the chips using the QuantStudio™ 3D Digital PCR Chip Loader 3 8. Transfer 12 μL of each DNA sample, diluted as appropriate, to the corresponding reaction tube. Mix well by gently pipetting up and down after each transfer (or gently vortex on low-medium speed). 9. Cap the reaction tubes, then briefly centrifuge them and immediately proceed to load the chips. IMPORTANT! For optimal results, load the chips as soon as possible after setting up the reactions. If you placed the reactions on ice, warm them to room temperature prior to loading. Load the chips using the QuantStudio™ 3D Digital PCR Chip Loader For instructions on loading chips manually, see Appendix E, “Load chips manually“. The following procedure describes how to load the QuantStudio™ 3D Digital PCR Chip v2 or QuantStudio™ 3D Digital PCR Chip automatically using a QuantStudio™ 3D Digital PCR Chip Loader. This procedure assumes that you have prepared your PCR reactions as explained in “Prepare the dPCR reaction mix“ on page 24. Before proceeding, see “Guidelines for loading and sealing chips“ on page 20. Materials required The following materials are required to load and seal chips. • Prepared digital PCR reactions • QuantStudio™ 3D Digital PCR Chip v2 or QuantStudio™ 3D Digital PCR Chip • QuantStudio™ 3D Digital PCR Chip Lid v2 or QuantStudio™ 3D Digital PCR Chip Lid • QuantStudio™ 3D Digital PCR Sample Loading Blades • Immersion Fluid syringe • Immersion Fluid syringe tip • QuantStudio™ 3D Digital PCR Chip Loader • UV-Activated Chip Sealant syringe (not required for v2 chips) • Isopropanol • Gloves, powder-free, nitrile • Scissors • Clean room-grade, low-lint polyester wipes • Microcentrifuge • Pipettes and tips, P10 to P1000 • Vortexer QuantStudio™ 3D Digital PCR System User Guide 25
