QX100™ Droplet Generator Instruction Manual Catalog #186-3002
Preface Bio-Rad Technical Support For help and technical advice, please contact the Bio-Rad Technical Support department. In the United States, the Technical Support department is open Monday–Friday, 5:00 AM–5:00 PM, Pacific time. Phone: 1-800-424-6723 Fax: 1-510-741-5802 Email: LSG_TechServ_US@bio-rad.com (for U.S. and international customers) Online technical support and worldwide contact information are available at www.consult.bio-rad.com.
Table of Contents QX100™ Droplet Generator 1.1 Introduction 1.2 QX100 Droplet Generator 1.3 Installation and General Operation 1 1 2 3 Droplet Generation 2.1 Sample Preparation 2.2 Operation of the QX100 Droplet Generator 2.3 Preparation for PCR 5 5 6 10 Specifications and Maintenance 3.1 Specifications 3.
Preface Safety and Regulatory Compliance This instrument has been tested and found to be in compliance with all applicable requirements of the following safety and electromagnetic standards: ■■ ■■ IEC 61010-1:2001 (2nd ed.), EN61010-1:2001 (2nd ed). Electrical Equipment for Measurement, Control, and Laboratory Use — Part 1: General requirements EN 61326-1:2006 (Class A). Electrical equipment for measurement, control, and laboratory use.
Preface PPE (Personal Protective Equipment) Training Proper use of gloves is recommended with use of oils and sample plates. OSHA requirements for PPE are set forth in the Code of Federal Regulations (CFR) at 29 CFR 1910.132 (General requirements); 29 CFR 1910.138 (Hand protection); 29 CFR 1926.95 (Criteria for standard personal protective equipment). Any gloves with impaired protective ability should be discarded and replaced.
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1 QX100™ Droplet Generator 1.1 Introduction The QX100™ Droplet Digital™ PCR (ddPCR™) system performs digital PCR with unrivaled accuracy and precision. The system consists of two instruments, the QX100 droplet generator and the QX100 droplet reader, and their associated consumables. The QX100 droplet generator partitions samples into 20,000 nanoliter-sized droplets, and after PCR on a thermal cycler, droplets from each sample are analyzed individually on the QX100 droplet reader.
Chapter 1 QX100 Droplet Generator 1.2 QX100 Droplet Generator The QX100 droplet generator uses microfluidics to combine oil and water (sample) to create the droplets required for ddPCR analysis. It processes up to eight samples at a time, in under 2 minutes. Following sample preparation using either the ddPCR supermix or One-Step RT-ddPCR kit for probes, 20 μl each of eight prepared samples (or blanks) and droplet generator oil are transferred to the droplet generator (DG™) cartridge.
Chapter 1 QX100 Droplet Generator Table 1.2. Additional materials required for droplet generation.
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2 Droplet Generation 2.1 Sample Preparation Prepare the PCR reaction by combining 2x PCR supermix, 20x primers and probe, and DNA sample. Mix by vortexing in short pulses, and centrifuge briefly. ■■ ■■ ■■ ■■ ■■ ■■ The concentration of intact human genomic DNA should be <66 ng per 20 µl reaction.
Chapter 2 Droplet Generation 2.2 Operation of the QX100™ Droplet Generator The QX100 droplet generator prepares droplets for eight samples at a time. Droplet generation takes 2 minutes for each set of eight samples (20–30 minutes for a 96-well plate).
Chapter 2 Droplet Generation 3. Transfer 20 μl of each prepared sample to the sample wells (middle row) of the DG8 cartridge. Air bubbles can cover the bottom of the well and result in 2,500–7,000 fewer droplets and poor data quality. They are difficult to see. To avoid creating air bubbles, use the following pipetting techniques, which also ensure samples wet the bottoms of the wells so they are wicked into the microchannels (necessary for proper droplet generation).
Chapter 2 Droplet Generation 5. Using a multichannel pipet, fill each oil well (bottom row) with 70 μl DG oil from the reagent trough. Filling the oil wells with droplet generator oil. 6. Hook the gasket over the cartridge holder using the holes on both sides. The gasket must be securely hooked on both ends of the holder; otherwise, sufficient pressure will not build up for droplet generation. Correct placement of the gasket over the cartridge holder. 7.
Chapter 2 Droplet Generation Press to open and close Status indicator llights QX100 droplet generator with DG8 cartridge in place. 9. When droplet generation is complete, all three indicator lights are solid green. Open the door and remove the holder (with DG8 cartridge still in place) from the unit. Remove the disposable gasket from the holder and discard. The top wells now contain droplets, and the middle and lower wells are nearly empty with a small amount of residual oil.
Chapter 2 Droplet Generation 2.3 Preparation for PCR 1. Pipet 40 μl of the contents of the top wells (the droplets) into a single column of a 96-well plate.
Chapter 2 Droplet Generation 2. Seal the PCR plate wells with foil immediately after transferring droplets to avoid evaporation. Use Easy Pierce foil plate seals (compatible with the needles in the QX100 droplet reader) and a compatible thermal plate sealer. a. Allow the plate sealer to warm up for at least 10 min before use. b. Place the foil seal over the plate and place the plate in the plate sealer. c.
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3 Specifications and Maintenance 3.1 Specifications 5 inches (127 mm) 11 inches (279 mm) 14 inches (356 mm) Weight 10 lbs. (4.
Chapter 3 Specifications and Maintenance 3.2 Maintenance Surfaces of the instrument may require general cleaning. Use deionized/distilled water for general wipe down with a slightly dampened cloth. For decontamination, 10% bleach followed by 70% ethanol and/or deionized/ distilled water may be used. Do not use acetone or tap water. Inspect equipment regularly for damaged external components or wiring. Do not use if damaged.
Appendix A Ordering Information QX100™ ddPCR™ System ddPCR Reagents Catalog # 186-3001 Description QX100™ Droplet Digital™ PCR System, includes droplet generator, droplet reader, laptop computer, software, associated component consumables 186-3010 ddPCR Supermix for Probes, 5 x 1 ml tubes 2x supermix, for use in sample preparation for droplet generator in the QX100 Droplet Digital PCR system 186-3026 ddPCR Supermix for Probes, 2 x 1 ml tubes QX100 Droplet Generator, includes droplet generator, 1 box
Appendix A Ordering Information Thermal Cyclers 185-1196 C1000 Touch™ Thermal Cycler with 96-Well Fast Reaction Module, includes C1000 Touch thermal cycler chassis, 96-well fast reaction module, USB flash drive 186-1096 T100™ Thermal Cycler, includes 96-well thermal cycler, power cord, tube support ring 16 | QX100 Droplet Generator Instruction Manual
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Bio-Rad Laboratories, Inc. Life Science Group 10026322 RevA A US/EG US/EG 10026321 Rev Web site www.bio-rad.
