Nippon Genetics FastGene qFYR User manual

Real-Time PCR System
FastGene®qFYR Real-Time PCR System
User Manual
Version 1.0
NIPPON Genetics EUROPE

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 2
Copyright & Disclaimer, NIPPON Genetics EUROPE GmbH
The copyright of this manual is owned by NIPPON Genetics EUROPE GmbH. No individual or
organization shall, in any form, copy, edit, publish, or translate the contents of this manual
(including but not limited to words, trademarks, logos, keystroke icons, lists, images, data, etc.)
into other languages without the authorization or written permission of the company. In violation
of the above statement, the Company will investigate its relevant legal liability.
NIPPON Genetics EUROPE GmbH strives to ensure that the contents of this manual are correct
when printed, but the company still has the right to modify the contents of this manual at any
time without prior notice. The contents of the manual shall be subject to the changed contents.
User manual version number: V1.0
User manual revision time: September 2022
Type of instrument applicable: FastGene® qFYR Real-Time PCR System
Software Release Number: V 1.0
Instrument production date: See production label

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 3
Introduction
Welcome to the FastGene®qFYR Real-Time PCR System. Please read this instruction carefully
so that you can use this instrument correctly. After careful reading, please keep it for future
inquiry when necessary.
Product information
Description: Real-Time PCR System
Product model: FastGene®qFYR Realtime PCR System
Software Name: FastGene®qFYR Analysis Studio
Software Release Number: V1
Product Size: 350 x 520 x 370 mm (length x wide x height)
Net Weight: 25 Kg

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 4
Enterprise information
Statement of intended use:
This product is used to run real-time fluorescence PCR experiments (Real-time PCR) and to
analyze the experimental data. The instrument is operated in a laboratory with the corresponding
reagents, to carry out rapid and accurate quantitative and qualitative detection of target nucleic
acids sequences from human and animal samples (such as blood, oral swabs, nasopharyngeal
swabs, body fluids, etc.) or other analytes, or to conduct melting curves, genotype analysis, etc.
Special statement:
Before installing and using this instrument for the first time, please read the instructions in this
manual carefully. Please take into consideration the possible scope of maloperation or non-
recommended usage and pay special attention to the possible consequences.
Intended Usage:
This product is for Research-Use-Only (RUO).
User requirements:
This product must be operated by trained professionals, who have studied this user manual.

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Table of Contents
Table of Contents 6
Chapter 1 Security and consideraons 10
1.1 Security Symbols and Markings 10
1.2 General Instrument Safety and Precauons 10
1.3 Personal Safety and Precauons 11
1.4 Electrical Safety and Precauons 11
1.5 Environmental Safety and Precauons  12
1.6 Biosafety and Precauons 13
1.7 Electromagnec Compability (EMC) Standards 13
Chapter 2 Product Overview 15
2.1 Scope of applicaon 15
2.1.1 Intended Use 15
2.1.2 Product Intended Use Environment 15
2.2 Applicaon Areas 15
2.3 Basic Principles  16
2.4 Main Funcons  16
2.5 Product Composion  16
2.6 Product Structure 17
2.7 Instrument Parameters and Characteriscs 19
2.7.1 Physical Specicaons 19
2.7.2 Instrument Technical Parameters 20
2.7.3 Soware Features 21
2.7.4 Chemistry Reagent Features 21
2.7.5 Consumables Specicaons 21
Chapter 3 Installaon and Transportaon of Instruments 22
3.1 Transport and Storage 22
3.2 Unboxing 22
3.3 Packing List 23
3.4 Installaon Requirements 23

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3.4.1 Instrument Working Environment Requirements 23
3.4.2 Computer Conguraon Requirements 24
3.5 Equipment Installaon  25
3.5.1 Equipment Appearance 25
3.5.2 Release Instrument Transport Lock 27
3.5.3 First Installaon and Run Protocol 28
3.6 Instrument Soware Installaon 29
3.6.1 FastGene® qFYR Analysis Studio Soware Installaon 29
3.7 Pre-preparaon for boot 32
3.7.1 Instrument Self-inspecon 32
3.7.2 System Start-up 32
3.7.3 Experimental Preparaon 33
Chapter 4 Operaon to Run the Experiment 34
4.1 Soware Start 34
4.2 Create New Experiment 36
4.2.1 New Experiment or Loading exisng les 36
4.3 Experiment Properes  37
4.3.1 Name 37
4.3.2 Plasc Type 37
4.3.3 Experiment type 38
4.3.4 Chemistry 39
4.4 Experiment Method 40
4.4.1 Reacon volume 40
4.4.2 Thermal lid temperature 40
4.4.3 Basic seng 40
4.5 Plate Setup 42
4.5.1 Setup 42
4.5.2 Quick Setup 44
4.5.3 Remove Wells from the Reacon 44
4.6 Run an Experiment 45
4.6.1 Insert a Plate or Reacon tubes 45
4.6.2 Run the Experimental Program 45
4.7 Save Experiment 47

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Chapter 5 Operaon of Dierent Experiment Types   48
5.1 Absolute Quancaon (Standard Curve) Experiment  48
5.1.1 Setup of a Standard curve     49
5.2 Experiment of Melng Curve     52
5.2.1 Setup of Melt Curve Analysis     52
5.3 Relave Quancaon Experiment（（（Cq（53
5.3.1 Setup of comparave Cq (（（Cq) Analysis   53
5.4 Genotyping Experiments     54
5.4.1 Set Up of a Genotyping analysis     54
5.4.2 Set Up of a High-Resoluon Melt Analysis (HRM)  55
5.5 Presence / Absence Experiments    56
5.5.1 Set Up of a Presence / Absence analysis    56
Chapter 6 Result Analysis and Data Export    57
6.1 Result Tab57
6.1.1 Buons in Toolbar      57
6.1.2 Other Toolbars       58
6.2 Result Overview      59
6.2.1 Amplicaon plot59
6.3.1 Plot Properes       59
6.2.2 Plot Sengs60
6.2.3 Threshold Sengs      60
6.2.4 Baseline Sengs      61
6.2.5 Target Sengs61
6.2.6 Results as Reacon Board     62
6.2.7 Results as List63
6.2.8 Raw Amplicaon Plot and Raw Melt Peak Plot   64
6.2.9 Standard Curve65
6.2.10 Melt Curve66
6.2.11 Gene Expression Map67
6.2.12 Analysis Sengs      68

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6.2.13 Genotyping Maps69
6.2.14 Raw Melt Curves of the High-Resoluon Melt analysis  70
6.2.15 Aligned Melt Curves of the High-Resoluon Melt analysis 71
6.2.16 Dierence plot of the High-Resoluon Melt analysis  72
6.2.17 Presence/Absence Plot 74
6.2.18 QC Summary       75
6.3 Data Export       77
6.3.1 Toolbars in Export Tab77
6.3.2 Data Export       77
6.3.3 Printed report79
Chapter 7 Instruments Maintenance     81
7.1 Instrument Cleaning      81
7.1.1 Cleaning the Instrument     82
7.1.2 Clean Sample-tray and Reacon-holes    82
7.2 Instrument maintenance     83
7.2.1 Keep Air-Circulaon      83
7.2.2 Keep Stable Electricity Supply83
7.2.3 Keep Clean       83
7.2.4 Waste Disposal       83
7.2.5 Fuse Replacement      84
7.2.6 Overheang Protecon      84
Chapter 8 Troubleshoong      85

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 10
Safety markings used on instruments
Symbol Heading Description
Caution
This symbol is used to indicate that non-compliance with
instructions or procedures may lead to physical injury or even
death or could cause damage to the instrument. Consult the
Operator's Manual.
Biohazard This symbol is used to indicate that certain precautions must
be taken when working with potentially infectious material.
Hot surface Indicates the presence of a hot surface or other high-
temperature hazard and to proceed with appropriate caution.
Electricity hazard Indicates the presence of high voltage at the instrument.
Appliance class 1
Indicates that insulation of voltage-carrying parts (= basic
insulation) and connection of touchable metal parts to the
protective conductor (= grounding) protects against excessive
touch voltages
CE symbol This symbol indicates European conformity.
Serial number Identification of the serial number of the instrument.
Date of
Manufacturing The date of manufacturing will appear next to this symbol.
Manufacturer The name and the address of the manufacturer will appear
next to this symbol.
Disposal
Do not dispose of this product as unsorted municipal waste.
Follow local municipal waste ordinances for proper disposal
provisions to reduce the environmental impact
of waste
electrical and electronic equipment (WEEE).
1.2 General Instrument Safety and Precautions
The operator shall not disassemble the instrument, replace components or debug
the instrument without the authorization of our company. If one of the above is needed, it must
previously be approved by our company, and completed by trained professionals, otherwise
unexpected consequences will occur.
Please avoid collision, damage the instrument and store carefully.
The user should immediately unplug the instrument from the power outlet and contact the
supplier or ask our professional maintenance staff if one of the following is to happen:
•The instrument is subjected to rain or flooding.
•Abnormal sound or smell in the instrument.
•Collision or damage to the enclosure.
•The function of the instrument changed.

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 11
It is forbidden to carry or move the instrument during the operation of the instrument.
The openings on this instrument are ventilated. In order to avoid excessive temperature, do not
block or cover these vents during the operation of the instrument, or use dust shields and any
other substances to cover the surface of the instrument.
If you need to install or transport this instrument, please contact our company to request
professional personnel or professional guidance. Otherwise, we will not bear any responsibility
for the damage caused by the instrument.
After the installation of the instrument, the transport lock should be properly retained in
accordance with the requirements of the technical personnel.
During the operation of the instrument, do not forcibly open the sample draw, otherwise it will
destroy the biological safety of the instrument, its electromagnetic radiation, and other protective
measures.
Do not force consumables (8 tubes / test tubes /96 well plates, etc.) not matched with this
instrument into the sample draw.
Failure to use this instrument in accordance with the manufacturer's prescribed method may
damage the protection provided by the instrument.
1.3 Personal Safety and Precautions
The weight of the instrument is heavy. Moving or lifting the instrument requires appropriate tools,
methods, or cooperation with others to complete. Please avoid moving the instrument alone
without the cooperation of others. Improper lifting of this instrument may cause physical injury,
pain and/or instrument damage.
Do not touch the power plug, power cord or switch with wet hands.
High surface temperature:
During the operation of the instrument, the heating module of the sample draw may become very
hot! During the experiment and after the experiment, do not touch the heating block directly with
your hand or any part of the body to avoid scalding. Please open the sample draw after the heating
block temperature drops to standby temperature and take out the sample.
1.4 Electrical Safety and Precautions
Prohibition
This instrument has a voltage harmful to the human body! At any time, before disassembling the
instrument shell, please cut off the instrument power supply. It is forbidden to replace
components when power lines are connected.

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 12
The shell of this instrument should be properly grounded through the power ground wire. Any
damage to the internal or external grounding path of the instrument may be dangerous.
Note: If leakage is found, please cut off the power immediately and stop using the instrument.
Note: Please cut off the power supply before moving the instrument.
Use the power cord provided with this instrument. If the original power cord is broken, worn, or
disconnected, the same power cord should be replaced as soon as possible.
In order to avoid the danger of electric shock, the power cord of this instrument must be reliably
grounded. The power cord of this instrument is a standard plug power cord. Please insert the
power plug into the appropriate three-wire grounding socket (the external network power supply
connected by the socket must be reliably grounded) to ensure the safe use of the instrument.
The electrical supply in which the instrument is located must be connected with ground wire.
Please carefully check whether the power connection is firm. Do not hard pull the power cord.
When the plug is plugged into the power supply, make sure the plug is fully plugged into the
socket.
Please keep the power plug and the power cable away from high temperature objects such as
heater. When using this instrument, please do not place any items on the power cord, do not put
the power cord in the place where people often walk.
The fuse type of this instrument is T10AH250VP, placed in the backup box at the power outlet
at the back of the instrument. The use of improper fuses may damage the instrument's line system
and cause fire. Before turning on the instrument power switch, check and make sure the fuse is
properly installed.
In order to prevent fire hazards, use this instrument only with the indicated fuse. When replacing
the fuse, please cut off the power supply, pull out the power cord, pry open the fuse box with a
word screwdriver, and replace the old fuse with the same new fuse.
1.5 Environmental Safety and Precautions
This instrument is only suitable for indoor use, indoor ventilation should be good, and free of
corrosive gas.
EDM may cause explosion hazard and is prohibiting the use of this instrument in the presence or
possible presence of flammable and explosive gases.
The working environment temperature of this instrument should be between 10~30and the
reate umdt soud 5%.
The working environment pressure of this instrument should be normal atmospheric pressure
(altitude less than 2000 meters).

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 13
1.6 Biosafety and Precautions
Biological hazard
The sample object of this instrument is nucleic acid. In practice, please regard it as a biological
sample with potential biological hazard. When handling and operating samples, generally
applicable safety precautions shall be taken and appropriate protective goggles, clothing and
gloves shall be worn.
Users are requested to comply with all applicable local or national regulations to complete the
disposal of waste samples and contaminated materials.
This instrument shall be treated as waste with biological pollution hazard. Before reuse, recycling,
or disposal, the instrument needs to be purified (including cleaning, disinfection and/or
sterilization). This instrument shall be handled in accordance with relevant local or local
regulations.
During any operation, if liquid samples overflow, appropriate disinfectants should be quickly used
for disinfection, to avoid the spread of pollutants to laboratory personnel or contaminated
instruments.
1.7 Electromagnetic Compatibility (EMC) Standards
This instrument should be installed and used according to the electromagnetic compatibility
information in this chapter.
The electromagnetic compatibility of this instrument is classified into categories A and groups I.
1. This instrument meets the emission and immunity requirements specified in the
EN 61326-1:2013 and EN 61326-2-6:2013
2. This instrument is designed and tested according to A kinds of instruments in the
CISPR 11:2016
3. Portable and mobile RF communication instruments may affect the use of this
instrument. It is recommended to stay away from or close the portable and mobile
RF communication instruments when using this instrument normally.
4. This instrument must be used with the power cord provided by our company, the
length of the power cord is not more than 1.5 m
Except the accessories provided by our company, the use of other manufacturer accessories may
lead to an increase in the emission of this instrument or a decrease in immunity.
1. Please evaluate the electromagnetic environment before this instrument is used.
2. Do not use this instrument next to strong radiation sources (such as unshielded
RF sources), otherwise it may interfere with the normal operation of the
instrument.

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 14
Note: It is the responsibility of the user to ensure the EMC environment of the instrument
so that the instrument can work properly.

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 15
Chapter 2 Product Overview
2.1 Scope of application
2.1.1 Intended Use
FastGene®qFYR Real-Time PCR Systems are used to run real-time fluorescence PCR
experiments (Real-time PCR) and to analyze the experimental data. The instruments are operated
in a laboratory, in conjunction with the corresponding reagents, for rapid and accurate
quantitative and qualitative detection of target nucleic acids from human samples (e.g., blood,
oral swabs, nasopharyngeal swabs, body fluids, etc.) or other analytes in the patient's body, or for
melting curves, genotype analysis.
2.1.2 Product Intended Use Environment
Institutions such as medical institutions, laboratories, and companies that meet the requirements
of the product operating environment.
1. diagnostic laboratory use
2. General laboratory research use. Its users are professional and technical personnel of the
hospital laboratory or general laboratory and need specialized training for PCR laboratory
technology and instrument, software operation, and have skilled relevant operation skills.
2.2 Application Areas
-Basic scientific research
-Clinical detection of pathogens
-Genetic screening
-Cancer diagnosis
-Diagnosis and treatment of cardiovascular diseases
-The drug genome
-Public Health and Safety Surveillance
-Food Hygiene and Quarantine
-Customs Import and Export Quarantine
-Transgenic and Pathogen Detection in Animals and Plants
-and others

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FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 16
2.3 Basic Principles
PCR principle is similar to the natural replication process of DNA. Its specificity depends on
oligonucleotide primers complementary to both ends of the target sequence. PCR consists of
three basic reaction steps: denaturation-annealing-extension. Repeated cyclic denaturation-
annealing-extending processes can amplify the target gene by millions of times in 1~2 hours, for
a high detection sensitivity.
The principle of real-time fluorescence PCR is to add a fluorescent reporter (probe or dye) to the
PCR reaction system. The target nucleic acid derived from DNA samples is qualitatively or
quantitatively detected by fluorescence detection system and real-time monitoring of the
fluorescence value. Real-time fluorescence PCR detection has higher specificity and sensitivity
than end-point PCR detection.
2.4 Main Functions
FastGene®qFYR Real-Time PCR Systems are special instruments for real-time fluorescence
detection. It integrates PCR amplification, fluorescence detection and data analysis. It can monitor
the increase of fluorescence in each test tube in real time as the amplification increases. After the
amplification, the software automatically processes the experimental data, carries on the
quantitative / qualitative, melting curve or genotyping analysis to the sample, displays and prints
the experimental results.
2.5 Product Composition
This product mainly consists of electronic control system, power module, temperature control
system, detection system, shell components and software(V1).

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 17
2.6 Product Structure
Diagram of the Structure of the Machine
1. Sample module2. Sample draw 3. Inlet 4. Transport lock
5. Power indicator 6. Status indicator 7. In/Out button 8. Vent
9. Power switch 10. power cord socket 11. USB interface 12. Fuse
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
12.

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 18
Schematic diagram of temperature control module
Schematic diagram of optical path module
Schematic illustration of optical scanning patterns

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 19
2.7 Instrument Parameters and Characteristics
2.7.1 Physical Specifications
Instrument specifications Size: 350 mm(L) × 520 mm (W) × 370 mm (H).
Weight󲷙25 kg
Packing specifications Size: 710 mm(L) × 540 mm (W) × 580 mm (H).
Weight󲷙36 kg
Power supply specification
power supply voltage
󲷙
AC 110-230 V
Frequency󲷙50 Hz.
Power󲷙750 w
Communication specifications USB ports
Environmental conditions
Temperature
󲷙
10~ 30.
Humidity󲷙5 H no condensation.
Atmospheric pressure󲷙85.0 kPa ~ 106.0 kPa.
Altitude󲷙< 2000m
Storage and transportation environment Temperature󲷙-20~ 55󲷋in the transport box.
Humidity󲷙5 H
Running noise Maximum noise when running instrument does not exceed 60dB（A）

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®  
FastGene®qFYR Real-Time PCR Systems Installation, Use, and Maintenance Guide 20
2.7.2 Instrument Technical Parameters
Thermal parameters
Heat cycle Peltier semiconductor
Temperature control mode Temperature control module
Maximum ramp rate 3.6 /s
Temperature accuracy ± 0.2 °C
Temperature uniformity ± 0.2 
Optical parameters
Excitation source Single color LED
Detection device High sensitivity MPPC（silicon photomultiplier tube）
Fluorescent channel number 4channels (2x FAM)
Applicable to fluorescent
Channel 1
FAM/SYBR Green etc.
Channel 2
VIC/JOE/HEX/TET etc.
Channel 3
ROX/Texas Red etc.
Channel 4
Empty
Channel 5
Cy5 etc.
Channel 6
FAM/SYBR Green etc.
Detection parameters
Sample size 1-96 wells
Sample volume 10-50 l
Repeatability of fluorescence
intensity detection V 
Detection of fluorescence
intensity V 5
Sample repeatability V  
Linear fluorescence intensity
detection A linear reresson oeffent r00 n 5 onentraton radents
Sample linear correlation A near reresson oeffent00 n 5 concentration gradients