DNA-Technology DTprime User manual

Operation Manual
DTprime
Real-Time PCR instrument
PART I
Operating Procedure
TS 9443-004-96301278-2010
DNA-Technology, Research & Production LLC
Protvino

2

3
Safety Information
IMPORTANT: READ THIS MANUAL CAREFULLY BEFORE USING THE INSTRUMENT
General Safety
Whenever see the sign Caution! Danger! refer to the manual.
Before using the “DTprime” Real-Time PCR instrument (hereafter –the “instrument” or
“device”), please read this section carefully and pay particular attention to the safety
information. To avoid accidents, as well as breakdown of the instrument and equipment used
therewith, observe the safety rules given below.
If safety rules are violated during exploitation or in case of unintended use of the instrument,
the protection provided by the instrument may worsen and a health hazard may arise.
Do not operate the instrument if humidity exceeds 80% in the room. Condensate formation
may result of a short circuit in the instrument electronics.
The instrument should be saved from shocks and falling.
The instrument must be stored and transported only in a vertical position.
After transport or storage in humid and /or cold conditions, dry out the instrument not less
than 3 hours before connecting it to the supply voltage.
Avoid any liquids or objects getting inside the instrument's cabinet. This may result in
breakdown of the instrument.
“DNA-Technology, Research & Production”, LLC bears no responsibility for any injuries or
damage to health caused by unintended use or unauthorized modifications of the instrument.
Electrical Safety
Prior to connecting the instrument to an electric grid, check the presence of protective
grounding in the socket to which the instrument will be connected as well as integrity of the
power cable. Connection to the socket without grounding is prohibited. Please use the
power cable provided in the set of delivery. If liquid is spilt inside the instrument, disconnect
it from the power supply immediately and contact local service center or distributor.
Please connect the instrument to an electric grid with the corresponding voltage, which is
mentioned on the identification plate of it.
Caution! Danger of electrical shock! Replacement of fuses of the instrument has
to be carried out in compliance with the rules and guidelines of electrical safety
regulations. Fuses have to be replaced only when the instrument is switched off. The
instrument is considered to be unplugged if it is unplugged of the electricity socket and the
USB cable is disconnected from the USB 2.0 port of a PC.
During Operation
Do not expose the instrument to heat and sunlight or bright light from other intensive light
sources.

4
Maintenance
Do not open the cabinet of the instrument! It does not contain user serviceable
components.
Note! Settings of motor controllers, calibration parameters of optical and temperature
modules of the instrument cannot be changed by the end user. Calibration of the instrument
has to be performed by the manufacturer in accordance with the internal regulations of quality
control. Calibration data, if necessary, can be provided by the service department of DNA-
Technology Research& production
Operational Safety
The instrument is designed to be used under the following environmental conditions:
indoors;
altitude up to 2 000 m;
temperature from 5 °C up to 35 °C;
maximum relative humidity of 80% for the temperature up to 25 °C;
main voltage from 100 up to 220 V, frequency 50/60 Hz.
The istrument is IVD medical equipment and it is intended for studing of DNA samples using
the polymerase chain reaction (PCR) method and the detection of the accumulated product in
real time (real-time PCR). The instrument does not carry a direct biological hazard. Servicing
and maintainance of the instrument should be performed by specially trained, qualified staff.
While operation with the instrument:
wear suitable protective clothing, gloves and secure eyes and face;
do not pipette solutions by mouth;
do not eat, drink, smoke or use cosmetic products in the operational area;
after handling samples and chemical reagents wash your hands carefully;
dispose of leftover / used reagents and waste in compliance with the regulations in
force;
read the instruction for use provided with the PCR kit before running the assay;
while running the assay, follow the instructions for use;
do not use a PCR kit after its expiration date;
use only the chemical reagents provided along with the kit and those, which are
recommended by the manufacturer;
do not mix chemical reagents from different PCR kits or lots.
Environmental Effect
Deinstallation of the instrument for maintenance or recycling purposes: if properly operated,
the device doesn’t bear biologically hazardous.
Recycling of the instrument should be carried out in accordance with the regulations in force
at the area. The device does not contain any substances, which may represent a direct threat to
the environment.

5
Notice
This instruction for use contains information protected by copyright. No part of it may be
reproduced without preliminary written consent of “DNA-Technology, Research &
Production”, LLC. Any software products, equipment and designations mentioned in the text
may represent trademarks of their holders.

6
Table of Contents
1. General Information ............................................................................................................................ 8
1.1. Intended Use and modifications of the instrument....................................................................... 8
1.2. Delivery Set.................................................................................................................................. 9
1.3 Technical specification.................................................................................................................. 9
1.4. Operating Conditions.................................................................................................................. 11
1.5. PC Requirements........................................................................................................................ 11
2. Instrument description and principle of operation............................................................................. 12
2.1. Fast Thermal Control System.....................................................................................................14
2.2. Optical system ............................................................................................................................ 14
2.3. Control and Display System....................................................................................................... 15
2.4. Marking. Symbols used for marking.......................................................................................... 16
3. Setting up the instrument................................................................................................................... 18
3.1. Instrument Unpack ..................................................................................................................... 18
3.1.1. Instrument Carrying............................................................................................................. 18
3.2. Instrument installation................................................................................................................ 18
3.3. Software Installation................................................................................................................... 19
3.3.1. Instrument Driver Installation.............................................................................................. 19
3.3.2. Installation of RealTime_PCR software.............................................................................. 25
4. Operating Procedure.......................................................................................................................... 29
4.1. Instrument Switching-on and RealTime_PCR Launch ......................................................... 29
4.2. Preparing the Instrument for PCR.............................................................................................. 30
4.2.1. Checkup of Geometrical Settings........................................................................................ 30
4.2.2. Optical Measurement Exposure Setting .............................................................................. 31
4.2.3. Checking the Cleanness of Thermal Plate Wells................................................................. 32
4.2.4. Tubes Height Measurement................................................................................................. 32
4.3 Creation of Plate protocol and Amplification Program............................................................... 33
4.3.1. Filling in of Plate protocol................................................................................................... 33
4.3.2. Creation/editing of Amplification Program......................................................................... 34
4.4. Placement of Tubes with Samples............................................................................................. 36
4.5. Startup of the amplification program.......................................................................................... 37
4.6 Amplification program running................................................................................................... 38
5. Maintenance ...................................................................................................................................... 40
5.1 General Guidelines...................................................................................................................... 40
5.2 Maintenance Procedure ............................................................................................................... 41
5.3 Disinfection requirements of the instrument prior to maintenance and repair procedures.......... 41
5.4 Troubleshooting........................................................................................................................... 42
6. Storage and shipment ........................................................................................................................42
7. Recycling........................................................................................................................................... 43
8. EMS declaration................................................................................................................................ 43
9. Warranties.......................................................................................................................................... 44
10. Packing Certificate .......................................................................................................................... 45
11. Acceptance certificate...................................................................................................................... 45
12. Cards of warranty repair.................................................................................................................. 47
13. List of claims and instrument disinfection procedure...................................................................... 49

7
Introduction
The “DTprime” Detecting Thermocycler (hereafter –the “instrument” or “device”) is
intended to conduct qualitative and quantitative PCR analysis dropping out electrophoresis
stage of PCR products in agarose gel when using PCR kits based on the fluorescent detection
principle. Measurement of the quantity of accumulated product of PCR amplification takes
place right in the course of reaction temperature cycles (real-time PCR). The qualitative
analysis is based on evaluation of kinetics of PCR visible part, carried out using relevant
mathematical tool.
The use of fluorescent detection of PCR products has a list of substantial advantages:
high detection specificity (if oligonucleotide probes are applied, which allow detecting
only of certain amplicons);
high efficiency;
reduced assay time;
carrying out detection in a closed tube, which practically eliminates contamination of
further experiments;
availability of quantitative assessment of initial DNA-matrix;
registering and accounting of the data in electronic format.
Fig.1. Instrument General View

8
1. General Information
1.1. Intended Use and modifications of the instrument
The “DTprime” real time PCR instrument is intended for carrying out an assay using
polymerase chain reaction (PCR) method and accumulated product detection in a real time
(real-time PCR). Detection of the accumulated PCR amplification product occurs directly
during the execution of the amplification program, which increases the informative value of
the study.
The field of application - clinical laboratory diagnostics in vitro;
Purpose of use –diagnostics;
Sample type - samples of human DNA and microorganisms extracted from samples of
biomaterial;
The instrument is manufacturing in the following modifications: 2М1, 2М3, 2М6, 4M1, 4М6,
5M1, 5M3, 5M6, 6М6, 2Х1 4X1, 5X1, 6Х1.
Modification of the instrument is defined by the following structure –XXX, where:
1) 1st X is figure from 4 to 5 and it indicates the number of optical detection channels;
2) 2nd X is the symbol M or X and it determines the thermal block format: M - 96 wells of
200 l, X - 384 wells of 45 l.
3) 3rd X is figure 1, 3 or 6 and it means a thermal block version such as 1 –monoblock; 3 or
6 - the number of thermal block sections.
The instrument is designed for simultaneous analysis of up to 96 or up to 384 samples
depending on the modification (M or X).
Preparation and initialization of experiments on the instrument is realized with an IBM-
compatible personal computer, and further implementation of a preset program may be
executed autonomously. On completing the amplification program, the instrument goes to the
storage mode, and if the option “Turn off device after the run” is activated on the window
“Running”, the instrument goes to the power standby mode.
The optical measurements results may preliminarily be evaluated during the execution of the
amplification program, whereas the final evaluation –at any time after completing the
program. All data of the last amplification are saved in the internal memory of the instrument
and may be read out even in the case of power failure during amplification. As soon as the
mains supply is restored, the instrument continues to function with full recovery of the
amplification program status. Neither failure of the operating system, nor disconnection of the
external computer will halt the experiment. Thus, the instrument is highly protected against
major factors effect, which may result in a failure during PCR implementation.
One computer may control operation of several instruments, including ones of different types
(DTlite, DTprime) connected to it via USB ports.
To operate the instrument, the RealTime_PCR software is used, which allows performing the
following operations:
Test creation and editing;
Plate protocol creation;
Checking geometrical and optical settings of the device;

9
Creation, launching, and monitoring of the amplification program;
Monitoring the execution of the amplification program
Analysis of optical measurements data.
Data analysis report.
Settings and diagnostics of the instrument.
Loading and unloading of analyzed samples may be automatic in case if the instrument is
built in a robotic system.
Instrument is open system and it is not depended of concrete PCR kits.
1.2. Delivery Set
No.
Description
Reference
Producer
Quantity
pcs.
1
DTPrime real time PCR instrument
RT96-01IN
DNA-
Technology
R&P
1
2
Instruction for use (Part I and Part II)
RT96-04IFU
DNA-
Technology
R&P
1
3
Computer communication cable USB 2.0
High Speed А-В
—
China
1
4
Power cable (10А, 250V, 3х0,75mm2)
—
Taiwan
1
5
Fuses. (10А, 5х20, 250 V)
179200.10
Germany
1
6
Distribution package of software
RealTime_PCR
version 1.0.0.198
and higher
DNA-
Technology
R&P
2
7
Packing unit
RT96-03PG
DNA-
Technology
R&P
1
The instrument may be delivered along with portable PC with the pre installed software in
accordance with customer’s request. Note: PC should be purchased along with the order of
the instrument.
1.3 Technical specification
Specification
Value
Thermal block format (modification M)
96 test tubes of 0.2 ml (12 x 8)

10
Thermal block format (modification X)
384-well microplate
Tube type (modification M)
0.2-ml test tubes for PCR (individual, in strips, 8
pieces each, or in plates 12 x 8)
Tube type (modification X)
384-well microplate, 24x16 wells of 45 mcl
volume
Reaction volume (modification M)
10-100 mcl (10-50 mcl recommended)
Reaction volume (modification X)
5-30 mcl
Temperature range
0 C-100°C
Temperature step
0.1°С
Temperature accuracy
±0.2°С
Temperature uniformity
±0.15°С
Average heating ramp rate within temperature
range of 0…100°С (modification M)
Maximum heating ramp rate within temperature
range of 0…100°С (modification M)
3.3°C/s
3.5°C/s
Average cooling rate within temperature range of
100...55°С (modification M)
Maximum cooling rate within temperature range of
100...55°С (modification M)
2.1°C/s
2.5°C/s
Average heating ramp rate within temperature
range of 0…100°С (modification X)
Maximum heating ramp rate within temperature
range of 0…100°С (modification X)
2.1°C/s
2.5°C/s
Average cooling rate within temperature range of
100...55°С (modification X)
Maximum cooling rate within temperature range of
100...55°С (modification X)
1.0 °С /s
1.5 °С /s
“Hot lid” temperature
105°С ±1°С
Heating and cooling method
Peltier elements
Excitation source
LED
Detector
CCD camera