This prevents you from losing a newly created method. You return to the top level screen and the method you created is not saved. Overview About This Chapter This chapter provides information about how to create and edit PCR methods and how to work with stored methods. You can add up to 19 different user names to the instrument. Use the User function on the Main menu to add new users or edit existing user names.
The name you add or the name you select from a list of existing user names becomes the current user name. All new methods that you create are stored by default under the current user name. To add a new user name: Step 1. Action From the Main menu, press User. The Select User Name screen appears. A list of names of all users who have been added to the instrument displays in a 4 x 5 matrix. Accept Backsp Cancel F1. Action In the User Name field, enter an alphanumeric name up to six characters in length.
You can take the following action:. Press Accept to accept a name. The Security Code screen appears. You can take the following action: If you Protecting Methods You can protect methods and prevent other users from accidentally overwriting or deleting them by entering a Personal Identification Number PIN on the Security Code screen. User Name: hank. Then you set protection to Locked to prevent methods from being overwritten or deleted. Accept Cancel F1.
Press Lock to lock your method. This toggles between a Locked and Unlocked state. The Protection field displays the status of the method. Press Accept when you have entered a PIN you want to keep. The new name you entered should now display on the screen. Press Cancel to cancel your entry and return to the previous screen. Changing a User If you know the personal identification number for a user name, you can use the Name Circular Key Pad to select that name, and change it.
To change a user name: Step 1. To delete a user name: Step. Press Delete to delete the name. This removes the name from the Select User Name screen and allows you to add a new name to the instrument. The create screen displays this default method. You can run the default method shown above, or use it as a template to create a customized method. For detailed information about these parameters refer to the table below: For information about Displaying the Follow the steps below to display the Create Methods screen.
Note The More function only displays when you select a time or temperature parameter. Control Parameters To enter temperature control parameters: Step. One 1 is the typical setting for most PCR amplifications and is the default value for the Hld field. Generally, pre-PCR holds define a temperature and hold-time sufficient to denature any endonucleases or exonucleases that may contaminate your prepared samples.
To define pre-PCR holds: Step 1. Note You can enter 0 in this field to delete the pre-PCR hold segment from your method.
Create the first temperature parameter: a. Press Enter. Type in a temperature value between 4. Create the first hold-time parameter: a. Type in a hold-time value between and minutes:seconds. Enter information for the next pre-PCR hold: a. Repeat step 2 and step 3 until you have hold-time and temperature values for each of the pre-PCR hold parameters you defined in step 1.
Action On the Create screen, in the Tmp field, type in the number of temperature control parameters 2—6 you want for the PCR segment of your method. Note Specifying only the minimum number of PCR cycles required for analysis will minimize the chance that unwanted targets will amplify competitively. Enter the number of cycles you want the method to run: a. Press Enter to select the Cycles field. In the Cycles field, type in the number of cycles from 2— Action Create the first hold-time parameter: a.
Repeat step 3 and step 4 until you have time and temperature values for each of the segment temperature control parameters you defined in step 1.
The post-PCR incubation temperature and hold time parameters define how to hold your samples at a specified temperature until you are ready to analyze them. Type in a hold time value between and min:sec. Repeat step 4 through step 7 until you have time and temperature values for each of the post-PCR hold parameters you defined in step 2.
Modifying Cycling Introduction In addition to customizing values for PCR temperature control parameters, you can use the More function on the create screen and access cycle modification functions that allow you to:. The time or temperature parameter you select on the create screen, determines which modification function you can access when you press More. Different modification functions are available depending on whether you select a temperature control parameter in the pre-PCR segment, the PCR segment, or the post-PCR segment of a method.
Changing Using the AutoX function, you can automatically increase or decrease the value for Temperature any PCR segment parameter by a fixed amount every cycle. Control Parameters Note This feature is particularly useful towards the end of the amplification process since the amount of PCR product, available to be extended, increases with the number of cycles while the amount of available enzyme remains constant. Note If you have inserted a programmed pause, the AutoX screen displays the pause, but you cannot modify it from the AutoX screen.
Select the PCR time or temperature parameter that you want modified when you run your method. Note From the AutoX screen, you cannot modify the number of parameters in each segment or the number of cycles. Modifying Ramp The ramp time is the time it takes the instrument to change from one temperature to Rates another.
To modify ramp rates: Step. In this example, you can modify the rate at which the instrument ramps up from Move the highlight box to the third temperature in the cycling segment of the default method to modify the highlighted ramp rate. The third ramp rate defines the rate at which the instrument increases from Note You can also use the numeric keys to enter a value that defines the percentage by which you want to decrease the ramp rate for each cycle of the method.
You can enter values between 5 and 95, or The asterisk remains beneath the modified temperature parameter to remind you that the method has been modified. Inserting Holds Use the Insert function to insert holds and cycles into your method, and program pauses that the instrument automatically inserts into your method as it runs. To insert holds: Step. From the Create screen, use the Circular Key Pad to select a time or temperature parameter to the right of where you want to insert a hold.
Press More. Depending on the parameter you select in step 1, one of three screens displays from which you can access the insert function. Note The Pause function will not display on the Insert screen if the highlighted segment already has a pre-programmed pause or if highlight box is not on a PCR segment parameter.
Press Hold to insert a hold of 4. From the Create screen, use the Circular Key Pad to select a time or temperature parameter to the right of where you want to insert a cycle. Inserting Follow the procedure below to insert a programmed pause. Programmed Pauses To insert a programmed pause: Step 1. Action From the Create screen, use the Circular Key Pad to select a PCR segment time or temperature parameter where you want to insert a programmed pause.
Define Programmed Pause Start first pause at cycle 1 of Pause every 25 cycles for Beep during the pause? No Accept F1. In the Start first Pause at Cycle field, type in the cycle number where you want the method to first pause.
In the Pause Every field, type in the pause frequency in cycles. The pause frequency specifies the number of cycles that will run between each pause. Press Accept to accept the pause information on the screen. The word Pause now displays to the right of the incubation step where you programmed the pause. Editing If you have inserted a programmed pause in your method, you can edit the Programmed Pauses parameters for the pause at any time.
To edit programmed pauses: Step. The Edit soft key appears. Press Edit to access the programmed pause screen. Naming and Storing Naming and storing completes the creation of the method. Methods To name and store methods: Step. Accept F1. Action In the Method Name field, follow these steps to enter a 1—16 character alphanumeric name. Step a. Action Use the Circular Key Pad to select a character in the list shown in the upper right portion of the screen.
After selecting a character, press Enter to place the character in the Method Name field. Press Accept after you have entered a method name. The Store screen appears again. The method name you entered should now display in the Method Name field. Printing a Method Introduction If you have configured your instrument for a printer, you can print a record of the parameters in a method. Action Access the Create or Edit screen. From the Create or Edit screen you can print a copy of the parameters for the method displayed on the screen.
Editing or Deleting Methods Introduction After you create a method, you can edit its parameters, and store the method by the same name, or change its name.
At some time, you may also want to delete a method if you are no longer using it. You can access all editing functions from the Main menu. The delete screen is accessed through the utility menu. Editing a Method The following procedure describes how to edit a method. To edit a method: Step 1. Action From the Main menu, press Edit.
The top line of the display continuously cycles between the following three lines: Methods on Inst [or on PC card]. Used Mem: xxx methods xxx segments Free Mem: xxx methods xxx segments The following table describes these fields.
Based on a calculation of the complexity and length of a method relative to a maximum size of size segments for the storage capacity of the instrument. Select one of the methods displayed on the screen, or select another method as follows: If you want to Edit temperature control or time parameters. Note Editing parameters on the Create screen involves the same tasks and uses the same key combinations as you use when creating a method.
The same functions for modifying methods are also available. Deleting a Method The following procedure describes how to delete a method. To delete a method: Step 1.
Action From the Utilities screen, press More and then press Delete. The Delete Method screen appears. Press Delete. The Delete Confirmation screen appears. If the method is protected, enter a four-digit PIN and press Accept when the number is correct. Action Press Yes to confirm the deletion. This deletes the method and returns you to the Delete screen. Note Even after you delete the last method stored under a User name, the name is removed from the instrument.
This information is based upon differences in how the instruments heat and cool samples. This is because the DNA Thermal Cycler and the DNA Thermal Cycler starts counting the hold time when the block reaches a temperature one degree before the target temperature, while the GeneAmp PCR System starts counting the hold time when the samples reach a temperature one degree before the target temperature.
Hold Time As shown in the figures below, since the block reaches the target temperature before Differences the sample, the programmed hold time on the DNA Thermal Cycler and the DNA Thermal Cycler must include enough time for the samples to reach the target temperature.
What to Base the The following table lists what to base the new hold time on if you are using an up ramp New Hold Times On or down ramp.
If you are using Determine the change in temperature required to reach the next target temperature and round this value off to the closest value found in Table Table PCR is a process in which millions of copies of a specific sequence of DNA can be made in a matter of only a few hours.
This is important for forensic DNA samples since the DNA often found at crime scenes is limited in both quantity and quality. This process takes approximately minutes to set-up and then approximately three hours to run on the thermalcycler instrument. After the PCR reaction is completed it results in a large mixture of amplified DNA molecules which need to be separated in order to distinguish the various molecules from one another.
This is accomplished through a process known as capillary electrophoresis. DNA molecules carry a negative charge and once an electric current is applied to the sample the molecules enter a very thin capillary filled with a gel-like polymer and migrate towards the positive anode at the other end of the capillary.
The data from this process is then collected on a computer attached to the CE instrument and then through the use of a software program a DNA profile is developed. There are many wonderful features in the e7 package, this manual will help you use them to your benefit. If more education is wanted, please contact your.
Micros vendor to purchase additional The following document contains a listing, by interface type, of all third party systems, which can be interfaced to the.
Oracle Hospitality Property Management Systems. This list Any system not listed will require testing and certification PDF version available for download at www. Jul 2, All Rights Reserved. Designed by Templatic. You can access all editing functions from the Main menu. The delete screen is accessed through the utility menu. Page 87 To edit a method: continued Step Action Press after selecting a method.
The Edit screen appears. To delete a method: Step Action From the screen, press and then press. Page 89 To delete a method: continued Step Action Press to confirm the deletion. This deletes the method and returns you to the Delete screen.
Note Even after you delete the last method stored under a User name, the name is removed from the instrument. This information is based upon differences in how the instruments heat and cool samples. Note The typical hold time is 10 to 15 seconds for denaturation. Page Up Ramp Example 35 seconds see the figure below. Add the indicated times instead of subtracting them. Page Routine Maintenance System In This Chapter All instruments have factory installed fuses.
However, if you should ever need to change a fuse, or if you need to convert from the factory configured single-line service to double-line service, follow the instructions that follow in this chapter. You need a Single-Line flat-tip screwdriver to open the door of the Power Entry Module and change the fuse.
Operation To change the fuse for single-line operation: Step Action Turn off the instrument and disconnect the power cord from the side of the Operation To change the fuses for double-line instruments: Step Action Turn off the instrument and disconnect the power cord from the side of the instrument. During a Power The following table lists the actions the instrument takes if the power is interrupted and Failure the instrument turns off while you are operating it.
Page The feature is not implemented in the current firmware Upgrade firmware when the version. Printer not responding The printer has been disconnected or is off line. Check printer connections and power switch. Page Troubleshooting Information Heated cover not responsive Heated cover failure.
Call Technical Support. Heating rate too slow Run Rate Test diagnostic. Peltier failure. Call Applied Biosystems Technical Support. Run Rate Test diagnostic. Page System Specifications System Specifications Dimensions The following tables list the footprint and the weight of the instrument with the 96—Well or 60—Well sample block module.
Footprint With Sample Block Module Installed Height 26 cm 10 in Width 28 cm 11 in Depth 41 cm 16 in Note You must provide sufficient space around the instrument for unrestricted air circulation.
Page Control Panel Specifications Control Panel Specifications Display Screen The display screen is a 7 x 40 character display with a graphics mode of 60 x pixel resolution. Keys The instrument control panel consists of a display screen and 22 keys. The keys are: Function keys Arrow keys Stop key Page Touchdown Pcr Touchdown PCR When the optimal annealing temperature is unknown, one strategy, touchdown PCR, incrementally decreases the annealing temperature in early cycles in order to maximize the yield of specific products.
Telephone or Fax Note To schedule a service call for other support needs, or in case of an emergency, dial North America 1. Page Follow the voice instructions to order documents for delivery by fax. Note There is a limit of five documents per fax request. Page Index Numerics cycling, modifying to AutoX function, using to DNA Thermal Cycler cycles, inserting converting hold times to deleting holds holds inserting alphanumeric characters, entering ramp rates, modifying to AmpliCycle Sequencing , B-2 about supplied method B-1, B-2, B-3 AmpliTaq Gold , B Page fire hazard GeneAmp PCR System instrument warnings Control Panel heated cover screen about heating, of samples specifications A-3 history of run, reviewing copying methods to history file records Page Create Methods screen, displaying parity bit , default method, about part numbers PCR holds, defining to for ordering accessories post-PCR holds, defining fuse part number pre-PCR holds, defining Pause Time Out, setting temperature control parameters, entering pauses, programmed editing or deleting methods to Page SRAM card, using to copy methods to starting instrument warning stop bit , chemical hazard Stop key, definition of chemical waste hazard storing methods to waste disposal system firmware, upgrading to Win 3.
Page Technical Support and Training appendix in this document. Print page 1 Print document pages. Rename the bookmark.
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