1.0 PURPOSE
To design a Standard Operating Procedure that describes the procedure for operation and maintenance of PCR machine.2.0 SCOPE
This Standard Operating Procedure (SOP) is applicable for the operation and maintenance calibration of PCR machine for in vitro studies
3.0 RESPONSIBILITY
3.1 All personnel in the invitro shall be responsible for implementation and compliance to this SOP.
3.2 HOD of Invitro shall be responsible to ensure the implementation and compliance to the SOP.
4.0 DISTRIBUTION
4.1 The Quality Assurance department responsible to keep SOP ‘Master Copy’ approved through Quality Manager. The copy of ‘Control Copy’ of SOP is being distributed in invitro department and placed near related Equipment/ Instrument as ‘Display Copy’.
5.0 DEFINITIONS
5.1 Definition(s)
5.1.1 PCR Machine also known as a thermocycler, is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR).
6.0 ABBREVIATIONS
6.1 Abbreviation(s)
6.1.1 SOP : Standard Operating Procedure
6.1.2 HOD : Head of the Department
6.1.3 PCR : Polymerase Chain reaction
6.1.4 DNA : Deoxyribose Nucleic Acid
6.1.5 UPS : Uninterrupted Power Supply
7.0 PROCEDURE
7.1 Instrument description
7.1.1 System Overview
7.1.2 PCR Machine has a thermal block with holes where tubes holding the reaction mixtures can be inserted. There is a controller part which controls the temperature values as per user’s settings. Controller part has display panel with touch screen for setting program and parameters.
Control Panel with Display
7.1.1.1 Touch screen controller display has buttons to set parameters
7.1.1.2 Detachable heating block has 96 tube holder to fir 96 well PCR plates and 0.2 ml PCR tubes
7.1.1.3 Lid has heating pad inside and knob outside for controlling better fit
7.1.1.4 Control Panel with Touch Screen Display
Control Panel with Command Options
7.1.1.5 For all the settings and command display panel has touch screen with buttons displayed
7.2 Operating the PCR Machine
7.2.1 Starting the PCR Machine
7.2.1.1 Make sure that power supply is on uninterrupted power supply (UPS)
7.2.1.2 Power on the system
7.2.1.3 Self test begins and login screen appears
7.2.1.4 Choose user and login accordingly
7.2.2 Using the PCR Machine
7.2.2.1 After Log In PCR machine home screen appears
7.2.2.2 Press <Quick Start> for the ser specific quick start programs
7.2.2.3 Press <Programs> to edit, save, copy or run programs
7.2.2.4 Press <Protocol Wizard> to open wizard tool
7.2.2.5 Press <Incubate> to incubate the sample block at a constant temperature
7.2.2.6 Press <Tools> to access the system settings, user management, documentation, backup tool or self-test functions
7.2.2.7 Press <Logout> to log out
7.2.3 Edit and Save Programs
7.2.3.1 Press <Programs> in the PCR Machine home screen. The program overview screen opens:
7.2.3.2 To edit a program template press <New form template>, select a template and press <Open template>
7.2.3.3 To edit a stored program select a user directory, select a program and press <Edit>.
7.2.3.4 Define the program name. Touch the input box “Name” and enter a program name using the PCR Machine keyboard.
7.2.3.5 Define the heated lid status and temperature. Touch the input box “Lid” and switch the heated lid on or off. If the heated lid is switched on enter a temperature between 30 °C and 110 °C.
7.2.3.6 To switch heated lid preheating mode on or off press the corresponding button.
7.2.3.7 Select a program step and press <Edit>. The edit step screen opens:
7.2.3.8 Use the cursor keys at the top of the screen to switch between program steps. The number of the currently activated step is displayed.
7.2.3.9 Touch the input box “Temperature” and enter the desired temperature in °C.
7.2.3.10 Touch the input box “Hold time” and enter the time <h:mm:ss>. To program a pause step press the button ∞.
7.2.3.11 To program loop touch the input box “Go to” and enter the step number the program shall go back to. Then touch the input box “Cycles” and enter how often the loop will be repeated.
7.2.3.12 To reduce or increase the annealing temperature from step to step within a loop press the button ∆T and set a negative or positive temperature increment.
7.2.3.13 To increase or reduce the incubation time from step to step in a loop press the button ∆t and set a time increment.
7.2.3.14 To adjust the ramping rate press the button ∆R and enter any value between 0.1 °C and max.
7.2.3.15 To define a gradient in gradient enabled units press the button Grad. In the next screen we can choose between standard <Std.> and linear gradient <Lin.> programming. When using standard gradient programming enter two temperatures for the first and last lane of the sample block. For linear programming enter the primer annealing temperature and define the temperature increment. The gradient curve and the temperatures for each lane are displayed. Press the button P to confirm the gradient settings.
7.2.3.16 To confirm all settings press the button P in the edit step screen. The software leaves the edit step screen and shows programming screen in graphical or spreadsheet view.
7.2.3.17 To insert a step select a program step in the programming screen and press <Insert>. The edit step screen opens. Enter the desired parameters and confirm the settings by pressing the button P.
7.2.3.18 To delete a step select a programs step in the programming screen and press <Delete>. In the security query confirm to delete the step by pressing the P button.
7.2.3.19 When the parameters for all steps are entered confirm the settings by pressing the button <Done> in the programming screen. The software opens the screen to copy and save programs.
7.2.3.20 To save the program select a user directory and a storage place, then press the button <Save>. Now the program is saved.
7.2.3.21 To copy programs press the button <Programs> in the PCR Machine home screen.
7.2.3.22 To copy all programs located in a user directory and press the button <Copy all>. Select the user directory the programs should be copied to and press the button <Save>.
7.2.3.23 To copy a single program select a user directory and a program and press the button <Copy>. Select a user directory, a storage place and press the button <Save>.
7.2.3.24 To delete all programs located in a user directory select a user directory, press the button <Delete all> and confirm the security query.
7.2.3.25 To delete a single program select a user directory and a program and press the button <Delete>. Confirm the security query to delete the selected program.
7.2.3.26 The PCR Machine thermal cycler stores the latest 5 edited or started programs for each user. For quick start of a program press the <Quick start> button in the PCR Machine home screen. Select a program from the list and press the <Start> button.
7.2.3.27 To start a program from a user directory press the button <Programs> in the PCR Machine home screen. In the next screen select a user directory and a program and press the <Start> button.
7.2.3.28 After start the running program is shown in spreadsheet, graphical or gradient view. Use the same buttons as for the programming screen is to switch between the different modes. Please note that the gradient curve can only be shown during a gradient step.
7.2.3.29 To pause the active program press the button <Pause>. The color of the highlighted step changes to blue and in the field “Step” the message Pause is shown.
7.2.3.30 To continue a paused program press the button <Continue>.
7.2.3.31 To skip a step press the button <Skip>. The program will continue with the next step.
7.2.3.32 To stop the active program press the button <Stop program> button and confirm the security query. Programs with a pause in the last step have to be stopped manually as well.
8.0 PRECAUTIONS
8.1 Protect the outer sleeves from damage, protect from sharp objects
8.2 Check gas level/pressure before beginning of the experiments
8.3 Only the power cord supplied with the workstation should be used to connect the workstation to the mains supply.
8.4 The exhaust valve outlet must not be covered or blocked.
8.5 The cooling fan covers and cooling vents must not be covered or blocked.
8.6 To minimize the effect of the pressure change, enter and exit the workstation chamber slowly
9.0 REFERENCE
8.1 Reference
8.1.1 PCR Machine Manual
END OF THE DOCUMENT
6.1 Abbreviation(s)
6.1.1 SOP : Standard Operating Procedure
6.1.2 HOD : Head of the Department
6.1.3 PCR : Polymerase Chain reaction
6.1.4 DNA : Deoxyribose Nucleic Acid
6.1.5 UPS : Uninterrupted Power Supply
7.0 PROCEDURE
7.1 Instrument description
7.1.1 System Overview
7.1.2 PCR Machine has a thermal block with holes where tubes holding the reaction mixtures can be inserted. There is a controller part which controls the temperature values as per user’s settings. Controller part has display panel with touch screen for setting program and parameters.
Control Panel with Display
7.1.1.1 Touch screen controller display has buttons to set parameters
7.1.1.2 Detachable heating block has 96 tube holder to fir 96 well PCR plates and 0.2 ml PCR tubes
7.1.1.3 Lid has heating pad inside and knob outside for controlling better fit
7.1.1.4 Control Panel with Touch Screen Display
Control Panel with Command Options
7.1.1.5 For all the settings and command display panel has touch screen with buttons displayed
7.2 Operating the PCR Machine
7.2.1 Starting the PCR Machine
7.2.1.1 Make sure that power supply is on uninterrupted power supply (UPS)
7.2.1.2 Power on the system
7.2.1.3 Self test begins and login screen appears
7.2.1.4 Choose user and login accordingly
7.2.2 Using the PCR Machine
7.2.2.1 After Log In PCR machine home screen appears
7.2.2.2 Press <Quick Start> for the ser specific quick start programs
7.2.2.3 Press <Programs> to edit, save, copy or run programs
7.2.2.4 Press <Protocol Wizard> to open wizard tool
7.2.2.5 Press <Incubate> to incubate the sample block at a constant temperature
7.2.2.6 Press <Tools> to access the system settings, user management, documentation, backup tool or self-test functions
7.2.2.7 Press <Logout> to log out
7.2.3 Edit and Save Programs
7.2.3.1 Press <Programs> in the PCR Machine home screen. The program overview screen opens:
7.2.3.2 To edit a program template press <New form template>, select a template and press <Open template>
7.2.3.3 To edit a stored program select a user directory, select a program and press <Edit>.
7.2.3.4 Define the program name. Touch the input box “Name” and enter a program name using the PCR Machine keyboard.
7.2.3.5 Define the heated lid status and temperature. Touch the input box “Lid” and switch the heated lid on or off. If the heated lid is switched on enter a temperature between 30 °C and 110 °C.
7.2.3.6 To switch heated lid preheating mode on or off press the corresponding button.
7.2.3.7 Select a program step and press <Edit>. The edit step screen opens:
7.2.3.8 Use the cursor keys at the top of the screen to switch between program steps. The number of the currently activated step is displayed.
7.2.3.9 Touch the input box “Temperature” and enter the desired temperature in °C.
7.2.3.10 Touch the input box “Hold time” and enter the time <h:mm:ss>. To program a pause step press the button ∞.
7.2.3.11 To program loop touch the input box “Go to” and enter the step number the program shall go back to. Then touch the input box “Cycles” and enter how often the loop will be repeated.
7.2.3.12 To reduce or increase the annealing temperature from step to step within a loop press the button ∆T and set a negative or positive temperature increment.
7.2.3.13 To increase or reduce the incubation time from step to step in a loop press the button ∆t and set a time increment.
7.2.3.14 To adjust the ramping rate press the button ∆R and enter any value between 0.1 °C and max.
7.2.3.15 To define a gradient in gradient enabled units press the button Grad. In the next screen we can choose between standard <Std.> and linear gradient <Lin.> programming. When using standard gradient programming enter two temperatures for the first and last lane of the sample block. For linear programming enter the primer annealing temperature and define the temperature increment. The gradient curve and the temperatures for each lane are displayed. Press the button P to confirm the gradient settings.
7.2.3.16 To confirm all settings press the button P in the edit step screen. The software leaves the edit step screen and shows programming screen in graphical or spreadsheet view.
7.2.3.17 To insert a step select a program step in the programming screen and press <Insert>. The edit step screen opens. Enter the desired parameters and confirm the settings by pressing the button P.
7.2.3.18 To delete a step select a programs step in the programming screen and press <Delete>. In the security query confirm to delete the step by pressing the P button.
7.2.3.19 When the parameters for all steps are entered confirm the settings by pressing the button <Done> in the programming screen. The software opens the screen to copy and save programs.
7.2.3.20 To save the program select a user directory and a storage place, then press the button <Save>. Now the program is saved.
7.2.3.21 To copy programs press the button <Programs> in the PCR Machine home screen.
7.2.3.22 To copy all programs located in a user directory and press the button <Copy all>. Select the user directory the programs should be copied to and press the button <Save>.
7.2.3.23 To copy a single program select a user directory and a program and press the button <Copy>. Select a user directory, a storage place and press the button <Save>.
7.2.3.24 To delete all programs located in a user directory select a user directory, press the button <Delete all> and confirm the security query.
7.2.3.25 To delete a single program select a user directory and a program and press the button <Delete>. Confirm the security query to delete the selected program.
7.2.3.26 The PCR Machine thermal cycler stores the latest 5 edited or started programs for each user. For quick start of a program press the <Quick start> button in the PCR Machine home screen. Select a program from the list and press the <Start> button.
7.2.3.27 To start a program from a user directory press the button <Programs> in the PCR Machine home screen. In the next screen select a user directory and a program and press the <Start> button.
7.2.3.28 After start the running program is shown in spreadsheet, graphical or gradient view. Use the same buttons as for the programming screen is to switch between the different modes. Please note that the gradient curve can only be shown during a gradient step.
7.2.3.29 To pause the active program press the button <Pause>. The color of the highlighted step changes to blue and in the field “Step” the message Pause is shown.
7.2.3.30 To continue a paused program press the button <Continue>.
7.2.3.31 To skip a step press the button <Skip>. The program will continue with the next step.
7.2.3.32 To stop the active program press the button <Stop program> button and confirm the security query. Programs with a pause in the last step have to be stopped manually as well.
8.0 PRECAUTIONS
8.1 Protect the outer sleeves from damage, protect from sharp objects
8.2 Check gas level/pressure before beginning of the experiments
8.3 Only the power cord supplied with the workstation should be used to connect the workstation to the mains supply.
8.4 The exhaust valve outlet must not be covered or blocked.
8.5 The cooling fan covers and cooling vents must not be covered or blocked.
8.6 To minimize the effect of the pressure change, enter and exit the workstation chamber slowly
9.0 REFERENCE
8.1 Reference
8.1.1 PCR Machine Manual
END OF THE DOCUMENT
You may like to read theses links:
1. List of All SOPs and Documents for Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for Animal House Facility
4. List of All SOPs and Documents for Clinical Research
5.List of All SOPs and Documents for Laboratory Instruments and Equipments
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment