Droplet Digital PCR

Preparing a GCR96 Cartridge for a
QX ONE™ Droplet Digital™ PCR System Run
Quick Start Guide

Transferring Droplet Digital PCR (ddPCR™) Reactions to a GCR96 Cartridge
Using a Rainin LTS P20 Multichannel Pipettor, transfer 20 µl of ddPCR reactions, consisting of RNA, DNA, or cDNA template, sample,supermix, assay(s), and nuclease-free water, into the wells of a GCR96 Cartridge that are designated by the labeled alpha-numericintersections (the wells that align with both a letter and number, as shown). Insert the pipet tips into the wells at approximately a 15° angle,
contacting the bottom of the wells. Dispense the ddPCR reactions slowly (approximately 5 sec). Do not push the pipet plunger past thefirst stop.
Note: Before transferring ddPCR reactions to a GCR96
Cartridge, thoroughly mix reagents after thawing them atroom temperature (vortex supermix for 30 sec after thawing).
Upon generating master mix(es) and after adding the sample/
template to the master mix, vortex once more (approximately
15 sec). Centrifuge the ddPCR reactions after vortexing tocollect the volume at the bottom of each well (30 sec, 1,150 rcf).
Centrifugation eliminates air bubbles and ensures that the ddPCR
reactions are at the bottom of each well. Mixing or vortexing is
critical to obtain homogeneity of reagents within droplets.

Note: GCR96 Cartridges contain six strips of 16 wells, asshown. Each strip of 16 is called a DG16 Cartridge. All DG16
Cartridges must be filled with ddPCR reactions or a 1:1 ratio ofddPCR Buffer Control for Probes (Bio-Rad™ Laboratories, Inc.,
catalog #1863052) and water. Failure to do so will affect theperformance of the QX ONE ddPCR System and subsequent
experimental data.
Sealing a GCR96 Cartridge
1 	Check the PX1 PCR Plate Sealer to ensure that the correct

settings are programmed for sealing the GCR96 Cartridge(Temp.: 180˚C; Time: 0.5 sec).
Alpha-numericintersection2 	
Securely load the GCR96 Cartridge on the silver heat block(Z-shaped support block; bottom protruding edge of heat
block should face the operator). Orient the GCR96 Cartridge
so that well A1 is at the upper left corner of the plate.
3 	Place the pierceable foil heat seal with the double red stripes

face up (facing toward the operator) on the GCR96 Cartridge.
Ensure the foil seal covers the entire surface area of thecartridge, with the exception of the cartridge labels identifying
rows A–H (left-hand side of cartridge) and the Droplet Digital
PCR label (right-hand side of cartridge), and that the seal
remains flush with the top and bottom of the cartridge (ensureno foil overhangs on any side).

Visit bio-rad.com/QXONE for more information.

Preparing a GCR96 Cartridge for a QX ONE Droplet Digital PCR System RunABCContact
betweenplate andbasket

eject the drawer. Load the heat block, cartridge, and seal intothe PX1 Sealer. Close the drawer by selecting Close in thetouch-screen menu on the PX1 Sealer (check to make sureseal is still aligned correctly before closing the drawer).
5 	Select Seal (Seal will be green when the PX1 Sealer has

reached the sealing temperature of 180˚C). The drawer willautomatically eject when the seal is completed.

BA

H

GFEDCBAABCDEFGH4 	
Select Eject from the PX1 PCR Plate Sealer touch screen toRefer to the QX ONE Droplet Digital PCR System and QX ONE
Software User Guide (10000116512) for additional details.
Visit bio-rad.com/QXONE for more information.
BIO-RAD, DDPCR, DROPLET DIGITAL, and QX ONE are trademarks of Bio-Rad
Laboratories, Inc. in certain jurisdictions. All trademarks used herein are the property of
their respective owner. © 2022 Bio-Rad Laboratories, Inc.

The QX ONE Droplet Digital PCR System, and the consumables and reagents designed
to work with this system, and/or their use is covered by claims of U.S. patents and/or pendingU.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc.
6 	
When the PX1 Sealer ejects the drawer, carefully rotate theSee bio-rad.com/en-us/trademarks for details. Purchase of the product includes a limited,
non-transferable right under such intellectual property for use of the product for internalsemisealed cartridge 180° (do not rotate the heat block fromresearch purposes in the field of digital PCR only. No
rightsare QXOne
granted Manual_Version1.1for diagnostic uses.
ProjectTitle:
its original orientation) and repeat steps 5–6. Then remove theNo rights are granted for use of the product for commercial
applicationsof any kind, including
Vendor: Troutt
Visual Services
but not limited to manufacturing, quality control, orFile
commercialservices, such as contract
sealed cartridge and heat block from the PX1 Sealer. StoreName: 11_centrifuge_basket_close_up
services or fee for services. Information concerninga license
for such uses can be obtainedDate:
08/26/20
the heat block on top of the PX1 Sealer until further use andfrom Bio-Rad Laboratories. It is the responsibility ofthe purchaser/end
user to acquire any
Proof:
1
Project Title: QXOne Manual_Version1.1
execute steps 7–9 with the sealed cartridge. Vendor: Troutt Visual Services
additional intellectual property rights that may be required.

Filerotor
Name: 11_centrifuge_basket
7 	Place the GCR96 Cartridge in a swinging bucket
Date: 08/26/20

centrifuge, placing row A closest to the rotor spindle
Proof: 1(refer toimages shown).
8 	Load a counterbalance cartridge on the opposite rotor bucket.
9 	
Push the individual DG16 Cartridges in the GCR96 frametoward the numbered edge as far as it will go to prevent
breaking the tabs of the cartridge. Centrifuge cartridges for30 sec at 1,150 rcf. Proceed with loading your cartridge into
the QX ONE ddPCR System. Refer to Initiating and Managing
a QX ONE Droplet Digital PCR System Run Quick Start Guide(bulletin 3431).

Bio-RadLaboratories, Inc.
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