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As shown in information on solubility

test, testing personnel can quickly

search for specific TM value for each

hole in the table.

Relative light intensity in solubility

analysis: As determined from the

absolute light intensity above,

fluorescence value variation trend is

clearly observed during solubility test.

The red line in the diagram will help

testing personnel in positioning the

temperature point with the most

intensive variation of fluorescence

strength.

Absolute light intensity in solubility

analysis: Where solubility test is

contained in initial test, the software

will draw absolute light intensity for

current solubility test, and the testing

personnel will visually observe

variation of fluorescence intensity

during test.

Description of solubility test

In QPCR test by dye method, solubility test is generally required. Test by dye

method has poor specificity, so solubility test will be performed to validate

whether primer dimer which is not concerns of the test is produced in test.

Содержание Real-Time PCR

Страница 1: ...Real Time PCR Detection System User Manual For X960A X960B ...

Страница 2: ...Section II Instrument Appearance and Internal Components P10 Overview P11 Construction Function Section III Instrument Description and Performance Properties P14 Instrument Description P15 Instrument Performance Indicators P16 Operating Environment and Storage Condition Section IV Use and Installation of Software P17 Installation Instructions P18 Computer Setting Win 7 P21 Start Installation P24 O...

Страница 3: ...to Instrument P34 New Project File P40 Specific Test Procedure P50 Test Considerations Section VI Other Function Introduction P52 Grade Calculation and Retrieval P53 Auxiliary TM Value Calculator P54 Logs Recording and Retrieval P55 Other Internal Operational Functions ...

Страница 4: ...ntenance of this equipment and provides instructions on proper installation use and maintenance of this instrument Safety Cautions Before installation use and maintenance of this instrument read and observe any cautions and considerations in this guide to ensure normal operation of the equipment and user s personal and property safety Introduction ...

Страница 5: ...ct shall be placed onto the power cable The cable shall not be located at area subject to busy traffic of persons Connect disconnect the power cable by holding the plug rather than any portion of the cable itself As high heat will be generated during operation of the instrument do not touch any metal module of the instrument with any part of body to prevent burn No object shall be located within a...

Страница 6: ...rsonnel in any of the following cases 1 Any reagent water or other liquid enters the instrument 2 The instrument falls down from an elevation or the enclosure is damaged 3 The instrument generates any abnormal noise or pungent smell during operation 4 Functions of the instrument vary substantially such as failure to start shut down or operate it normally ...

Страница 7: ...d the adhesive tape sealing the case Take out the accessories of the case and store them in a secure place for future use Remove EPE foam covered on the instrument and lift it up by supporting its bottom with two persons and then locate it on a proper lab desk Handle the instrument with caution Store the packing case properly for containing in future Section I Instrument Installation ...

Страница 8: ...cause ache in or permanent injury to body or fall of the instrument Lift or move the instrument with proper handling tool or in a correct way Two persons must be used to lift the instrument up This instrument shall be mounted upright on a horizontal surface around which there shall not be any vibration EMI or any high inductance equipment e g refrigerator centrifuge and stirrer ...

Страница 9: ...erly Do not touch the power cable with wet hand Do not disconnect the cable during operation of the instrument In case of any damage in the power cable replace it with an acceptable one The instrument shall be always connected to a socket earthed properly When the power supply is disconnected connect one end of the network cable provided to the LAN port at the back of the instrument and the other ...

Страница 10: ...Connection of Instrument Connect laptop to the instrument Connect PC to the instrument ...

Страница 11: ... Lid Handle USB Port Pad Tray Operation Indicator LED Power Switch Power Socket Fuse Air Outlet 232 Interface LAN Port WAN Port Air Outlet Foot Pad Air Inlet Section II Instrument Appearance and Internal Components ...

Страница 12: ...g The handle provided on the instrument is intended for push pull Do not lift up or press down the handler with force to prevent damage to the lid structure When the knob is switched to the point UNLOCK the lid can be slide back and forth for loading unloading of test tube After loading the test tube close the lid and rotate the knob clockwise to a proper position and run the program for testing I...

Страница 13: ...he tray For this instrument the tray moves in a range between 0 90 to the enclosure do not move the tray out of this range to prevent damage to the instrument There is a status indicator LED under the front panel which turns on in any of three colors blue green and red respectively indicating Standby Operating and Alarm ...

Страница 14: ...nlet to prevent it from being absorbed in and thereby permanent damage to the fan or cooler and damage to the instrument resulted from overheating of module component due to poor ventilation Do not place any object at any of the air outlets to prevent overheating and thereby permanent damage to the instrument resulted from poor ventilation Do not place any inflammable explosive or heat sensitive m...

Страница 15: ...onstruction It consists of power module control module driver module display module heated lid module sample base module camera module light source module and optical fiber module Application Range It is applicable to polymerase chain reaction PCR Section III Instrument Description and Performance Properties ...

Страница 16: ... holes x 0 2ml 96 holes x 0 2ml Light Source Ultrahigh luminance mono color LED Ultrahigh luminance mono color LED Detector High sensitivity refrigerator CCD High sensitivity refrigerator CCD Dynamic Testing Range 100 1010 100 1010 Minimum Testing Template Single copy Single copy Reaction Volume 15μl 100μl 15μl 100μl Fluorescence Excitation Wavelength 470nm 525nm 470nm 625nm Fluorescence Detection...

Страница 17: ...mi conductor thermoelectric module Temperature Control Programming Max 999 cycles Max 999 cycles Special Functions Threshold analysis dissolution curve analysis grade and customized parameters Operating Environment and Storage Condition Operating Environment Ambient temperature 10 30 Relative humidity 70 Storage Condition Ambient temperature 20 55 Relative humidity 80 To ensure Normal Operation of...

Страница 18: ...h shall be ignored if occurring Shut down any firewall installed in the computer Installation Considerations Note To avoid failure of installation resulted from any error occurring during installation and facilitate your installation please observe the following procedure Before Start Note Before installation of the software set your computer properly to prevent interruption of test and thereby da...

Страница 19: ...Computer Setting Win 7 Display Setting 1 Navigate to Start Control Panel 2 Select Power Options in Control Panel 3 Select Change when the computer sleeps in the window ...

Страница 20: ...4 Select Never under Turn off the display in the window Select Never under Put the computer to sleep 5 Return to the window in Step 3 and click on Personalization button ...

Страница 21: ...6 Click on Screen Saver at the lower right corner 7 Select None from the drop down list in the dialog box Single click ...

Страница 22: ...e a destination folder to install the program Step 3 Read the pre installation instructions Step 2 Double click on the setup file and choose a proper installation language Step 1 Access the setup package from the flash disk provided ...

Страница 23: ...Step 8 Install the program Step 7 Confirm any information before installation Step 6 Choose whether to create a shortcut on desktop Step 5 Choose name of the folder to be created under start menu ...

Страница 24: ... on Yes for automatic connection Step 10 Open page of RealGen Step 9 Finish the setup wizard and check whether RealGen is running check the option and click on Finish If the instrument connection fails check whether it has been properly connected to the computer if yes shut down any anti virus software and restart the instrument and then try connection after it beeps If connection fails again cont...

Страница 25: ...oad Project Load a finished project file to start a test Export Project Export a loaded project for data storage Import Standard Curve Import a standard curve with similar circumstance for analysis Running Logs Display any projects running currently in the program Print Print desired information Patient Information Import patient information for printing and data analysis ...

Страница 26: ...onitoring page Analyze Switch to subsequent analysis page note this makes sense only after a test file has been loaded Toolbar Project Change parameters After a test has been loaded when current settings are not proper to the expectation they may be changed under this option Any change will be applied immediately after completion Change to parameters makes sense only after a test has been loaded P...

Страница 27: ...roject in it and operate or modify a project Open Test File Open a test file list and manage any test file and load or delete a file Instrument List Open an instrument list and connect or rename any instrument in it Software Version Display current software version number Current Address Display IP address of the computer ...

Страница 28: ...pecify a reviewer for the project and save it by pressing ENTER This option can be left blank Password Specify a password to access this project It can be left blank Confirm the password Confirm the password by repeating it Test Result Path The path where the test result is saved Project File Path The path where the project file is saved Test Type Specify the type of current test ...

Страница 29: ...ired for current test At least one channel is required Cross Interference Call cross interference setting page to compensate interference between multi channel tests General Settings Call general setting page to perform some basic settings ...

Страница 30: ... used along with sample concentration Negative Negative comparison type Positive Positive comparison type Unknown Sample type in unknown concentration Gap Gap comparison type where water is generally used for comparison Unused No sample is used for current hole position Sample Concentration Specify sample concentration for standard sample Concentration Multiply Ascending Descending Quickly mark ho...

Страница 31: ...ition Sample Name Identify the sample for the hole position Housekeeping Gene Identify housekeeping gene for current test Standard Sample Identify standard sample in contrast group for current test Project Group Divide projects into different groups ...

Страница 32: ...program setting file edited currently Low Temperature Storage For storage at low temperature upon completion of test Solution Curve Enable solution curve function Save Save current project file upon completion of test Operate Operate any project file saved when the instrument is connected and idle ...

Страница 33: ...Analysis Interface Introduction It is used for analysis of test result after completion of test including baseline setting threshold analysis outcome analysis solubility curve and quantitative analysis ...

Страница 34: ...he instrument on and connect it to a PC by cabled or wireless means Open the program when the instrument beeps and the following information should be displayed If not refresh the instrument list by using right button Step 2 Click on Yes to connect it directly or manually connect it in toolbar ...

Страница 35: ...New Project File Click on New Project in project wizard dialog box Enter basic setting page Choose and enter proper settings and then click on Next ...

Страница 36: ......

Страница 37: ...quantitative test hole position must be marked or otherwise standard curve will be unavailable and there are at least three standard samples in gradient You may import any existing hole setting file Quickly mark hole positions arranging in rows or columns with concentration multiply ascending descending ...

Страница 38: ...ve test gene and sample for each desired hole shall be identified and housekeeping gene and reference sample shall be marked for the test When necessary project groups can be identified for distinction Upon completion click on Next ...

Страница 39: ...the temperature point chosen Set the selected temperature point as cycle end point Set the selected temperature point as cycle start point Delete a temperature point chosen Change cycle number Change or specify cycle number by dragging from a temperature point to an end point ...

Страница 40: ... in gradient Time Variation Increase time variation for each cycle which will be reflected to the point in each cycle Heating Rate Change heating rate and the default is Fastest Cooling Rate Change cooling rate and the default is Fastest Collect Fluorescence Mark it to collect fluorescence at current temperature point Only one temperature point can be specified for each test Set it by referring to...

Страница 41: ...t Set whether low temperature storage is enabled and the storage temperature This option is disabled by default Solubility Setting It is recommended to use the default value It is used to enable or disable solubility test which is settable in customization Save the project Import existing temperature control program export a compiled temperature control program ...

Страница 42: ...nd the range of linearity in logarithmic phase analysis is 0 1011 copies ml Generally the number of pathogens in a clinical specimen is 0 1010 copy within which quantitative PCR will dose accurately and the specimen need not be diluted Repeatability Quantitative PCR presents a stable result since the threshold value is set in exponential amplification phase during which concentration of the reacti...

Страница 43: ...d and then choose the target program and right click on Run After being started the software module will display module information lid information and estimated time After being started the software module will display the increasing temperature for current project After being started the software module will display the increasing temperature curve for current project ...

Страница 44: ...rrent project When solubility curve is contained in the project the software module will display current temperature variation When solubility curve is contained in the project the software module will display current fluorescence variation As necessary you can selectively review channels and holes ...

Страница 45: ...ameters Upon completion of test the test file will be automatically saved to the directory as set for the project Test results are classified into result with CT value absolute quantitative result with standard curve relative quantitative result with histogram SNP result with three pile diagram and HRM result with differential diagram ...

Страница 46: ...nd perform real time monitor of the whole PCR process by using fluorescence signals and carry on quantitative analysis on the unknown template through s standard curve During operation user can perform manual settings based reagent property in Reagent Parameters Setting at the right side including agent property sample concentration concentration multiple channel gene name marking sample name mark...

Страница 47: ...achieve a more suitable CT value In analysis interface you can choose different channels and make analysis by manually setting baseline or review curves of a certain type Baseline Baseline is a straight line along which fluorescence signals vary slightly in the first several cycles of PCR amplification reactions ...

Страница 48: ...ssa represents logarithm of initial copies and the ordinate indicates CT value Therefore initial copies for an unknown sample can be calculated from the standard curve only by determining CT value of the sample In this table you can view CT and TM values of a certain curve and other result information Fluorescence Threshold Setting Generally fluorescence signals in the first 15 cycles in PCR react...

Страница 49: ... be imported or exported in the interface Parameters Related to Standard Curve Standard samples are required in real time absolute quantitative PCR test The sample used to create standard curve can be in particle Determination of molecular weight of standard sample Concentration of DNA sample is 50 μg ml when OD260 value is 1 based on the following formula Concentration of sample particle μg μl OD...

Страница 50: ...are specified in test analysis and housekeeping gene and standard samples have been set Description of relative quantitative test In relative quantitative test gene and sample names need to be specified and housekeeping gene and standard samples need to be marked In lab test this method is mostly used in final test analysis Where error analysis is required each gene and sample need to be re tested...

Страница 51: ... point with the most intensive variation of fluorescence strength Absolute light intensity in solubility analysis Where solubility test is contained in initial test the software will draw absolute light intensity for current solubility test and the testing personnel will visually observe variation of fluorescence intensity during test Description of solubility test In QPCR test by dye method solub...

Страница 52: ...ons issued by the authorities shall be complied with by the laboratory Fluorescent material free disposable gloves disposable dedicated centrifugal tube self removing pipette and suction head with filter element shall be used Prepare and handle reaction fluid in such way to avoid generation of air bubble check whether reaction tube is sealed tightly prior to startup to prevent leakage of fluoresce...

Страница 53: ...lization with other waste The test platform and all test appliances shall be disinfected with sodium hypochlorite of 1 alcohol of 75 or UV lamp PCR reaction mixed liquor shall be stored at low temperature and protected from sunlight The test platform shall be cleaned after completion of test to avoid contamination ...

Страница 54: ...rade from main menu to access View Grade page For convenience of user the software provides a grade table for user s retrieval Upon entry of central temperature and gradient width the software will automatically calculate specific temperature value for each column and helps to ensure accurate test result ...

Страница 55: ... TM Value Calculation Click on TM from main menu to access TM Calculation page Enter a value and then click on Calculate to output the result The program is furnished with an embedded TM value calculator for user s access when necessary ...

Страница 56: ...ding and Retrieval About Logs Click on Logs from the main menu to access Logs page and view test operation logs Facilitate management of test history records by user and retrieval of related test history records ...

Страница 57: ...ons Chinese English Toggle Button Click on File Load Test in main menu Load test data In the popup dialog box select the target data in format of jgtest stored previously to load Click on Tool Chinese or English to change the language ...

Страница 58: ...a temperature segment to modify a parameter in the popup dialog box Real Time Modification to Operation Parameters Modification can be made only by double clicking on a temperature segment currently not in operation or otherwise a message box will occurs indicating No modification can be made to any node currently in operation ...

Страница 59: ...ic Instrument Company Limited Address Floor 4 Building 1 16 the Third Xiyuan Road Sandun Town Xihu District Hangzhou Zhejiang Province Telephone 0571 89905515 Facsimile 0571 89905515 802 Postal Code 310000 Email shanghai healforce com Web www healforce com ...

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