To Next PageTo Next PageWhy am I getting extremely high ?R or n R values with Applied Biosystems Laboratory Equipment?
Extremely high ?R or n R values with Applied Biosystems Laboratory Equipment can be due to a couple of reasons. First, ensure that ROX dye was selected as the passive reference when setting up the plate document; if not, select it. Also, check for evaporation by making sure that the reaction plate is sealed completely, especially around the edges.
Why is there high variability across the reaction plate with Applied Biosystems Laboratory Equipment?
High variability across the reaction plate when using Applied Biosystems Laboratory Equipment can be caused by a couple of factors. First, ensure that ROX dye was selected as the passive reference when setting up the plate document; if not, select it. Evaporation can also lead to variability; make sure that the reaction plate is sealed completely, especially around the edges, and use reagents that contain ROX passive reference dye.
What to do if I have low ?R or R values with Applied Biosystems 7500?
Low ?R or R values with Applied Biosystems Laboratory Equipment can occur due to a couple of reasons. If the extension time is too short, use the default thermal profile settings. Primer-dimer formation can also cause this; to ensure optimal results, run the reaction plate as soon as possible after completing the reaction setup. If you cannot run a reaction plate within 2 hours after completing the reaction setup, refrigerate or freeze the reaction plate until you can load and run it on the 7500 Fast instrument.
Why does the Applied Biosystems Laboratory Equipment run take more than 40 minutes?
If a run with Applied Biosystems Laboratory Equipment takes more than 40 minutes, it is likely because the thermal cycler mode is set to Standard or 9600 Emulation. Ensure that the thermal cycler mode is set to Fast.
What causes high variability across replicates in Applied Biosystems 7500?
High variability across replicates in Applied Biosystems Laboratory Equipment can be caused by the reaction mix not being mixed well. To resolve this, mix the reaction mix gently by inversion, then centrifuge briefly before aliquoting to the reaction plate.
What to do if the Rn vs. Cycle plot is not displayed in Applied Biosystems Laboratory Equipment?
If the Rn vs. Cycle plot is not displayed when using Applied Biosystems Laboratory Equipment, it is likely because ROX dye was not selected as the passive reference when the plate document was set up. To fix this, select ROX dye as the passive reference when you set up the plate document.
General| Detection Method | Fluorescence |
|---|---|
| Sample Capacity | 96-well plate |
| Thermal Cycler | Yes |
| Temperature Uniformity | ±0.5°C |
| Type | Real-Time PCR System |
| Thermal Accuracy | ±0.25°C |
| Detection Channels | 5 |
| Weight | 45 kg |
| Power Requirements | 100-240 V, 50/60 Hz |
| Dynamic Range | 9 logs |
| Reaction Volume Range | 10-100 µL |
| Temperature Range | 4°C to 100°C |
| Thermal Cycler Block | 96-well |
| Software | SDS Software |
| Detection | Multiplex (up to 5 colors) |
Initial computer setup before system installation.
Choosing between the setup wizard or manual guide.
Outlines the workflow and time for system installation.
Guide on using the automated setup wizard for installation.
How to follow the manual for manual system installation.
Prerequisites and materials needed before starting instrument setup.
Guidance on choosing optional hardware and software for the system.
Procedures for safely removing and positioning the instrument.
Verifying all components were received with the instrument.
Connecting and preparing the instrument for operation.
Network connectivity considerations for the system.
Recommendations for backing up system data to prevent loss.
Guidelines for installing additional software on the system computer.
Considerations for firewalls and encryption software compatibility.
Recommendations for system utilities like defragmentation.
Detailed steps for positioning the instrument crate and its contents.
Introduction of software installation and power-on process.
Step-by-step guide to install the SDS software.
Procedures for turning on the instrument after software installation.
Verifying instrument functionality after setup.
Common issues and solutions during installation.
Diagnosing issues based on instrument front panel indicators.
Diagnosing issues from instrument function test results.
Steps to prepare the calibration plate for ROI calibration.
Creating the necessary plate document in the SDS software.
Executing the ROI calibration procedure.
Common problems encountered during ROI calibration.
Introduction to the ROI calibration process.
Explains why ROI calibration is necessary for the system.
Instructions for loading the prepared plate into the instrument.
Procedures for removing the plate after calibration.
Resolving issues related to faint or absent ROI images.
Steps to prepare the background calibration plate.
Creating the plate document for background calibration.
Executing the background calibration procedure.
Analyzing the data obtained from background calibration.
Performing optical calibration specific to 7500/7500 Fast systems.
Common problems during background and optical calibration.
Introduction to background and optical calibration.
Explanation of background fluorescence in the system.
Best practices for performing calibrations.
Procedures for removing the background plate.
Performing optical calibration specific to 7500/7500 Fast systems.
Creating the plate document for optical calibration.
Analyzing data from the optical calibration run.
Procedures for removing the plate after optical calibration.
Resolving issues during background calibration.
Steps to prepare for pure dye calibration.
Creating the plate document for pure dye calibration.
Preparing and running the pure dye calibration plates.
Analyzing data from the pure dye calibration.
Evaluating the spectral data for pure dye calibration.
Introduction to the pure dye calibration process.
Explains the purpose and function of pure dye calibration.
Lists the dye sets supported by the instrument.
Information on using custom dyes with the system.
Analyzing data from the pure dye calibration.
Steps to prepare the RNase P verification plate.
Creating the plate document for RNase P verification.
Executing the RNase P verification run.
Analyzing data from the RNase P verification run.
Common problems during RNase P verification.
Introduction to instrument performance verification.
Explains the purpose of RNase P verification runs.
Steps to prepare the RNase P verification plate.
Creating the plate document for RNase P verification.
Executing the RNase P verification run.
Analyzing data from the RNase P verification run.
Procedures for removing the RNase P plate.
Resolving issues with RNase P plate data or run failures.
Procedure for shutting down the instrument for short periods (<7 days).
Procedure for shutting down the instrument for extended periods (>7 days).
Guidelines for weekly, monthly, and semi-annual maintenance tasks.
Procedures for managing and backing up system data files.
How to clean the sample block to remove contaminants.
Steps to defragment the computer's hard drive.
Guidelines for safely moving the instrument.
Checking the status of the instrument's halogen lamp.
Procedure for replacing the halogen lamp.
Procedure for replacing instrument fuses.
Guidance on operating system updates.
Maintenance tasks performed every six months.
Tasks for resolving specific problems as they arise.
Steps for cleaning and defragmenting the computer's hard drive.
Guidelines for safely moving the instrument.
Checking the status of the instrument's halogen lamp.
Procedure for replacing the halogen lamp.
Procedure for replacing instrument fuses.
Guidance on operating system updates.
Introduction to creating a custom background plate.
Step-by-step instructions for creating a background plate.
Introduction to creating custom pure dye plates.
List of materials needed for creating custom dye plates.
Steps to create a custom pure dye plate.
How to determine the optimal concentration for custom dyes.
How to add custom dye information to the SDS software.
Unpacking computer hardware and setting voltage.
Connecting computer peripherals and powering them on.
Configuring computer display and power settings.
Network connection and software download steps.
