Why am I getting extremely high ?R or n R values with Applied Biosystems 7300?
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 7300?
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 7300?
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 7300 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 Laboratory Equipment?
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 7300?
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.
Why do I get high C values or failed PCR reactions with Applied Biosystems 7300?
High C values, poor T precision, or failed PCR reactions when using Applied Biosystems Laboratory Equipment can occur due to several reasons. It could be that the target is difficult to amplify; in this case, increase the annealing/extension time in the thermal cycler protocol or increase the annealing/extension temperature to 62 °C. It may also be due to insufficient cDNA template; use 10 to 100 ng of cDNA template per 20-µL reaction. Poor cDNA template quality can also cause this. Quantify the amount of cDNA template and test it for PCR inhibitors. Sample degradation is another potential cause, so prepare fresh cDNA and repeat the experiment. Ensure that you are using the correct Master Mix, specifically the TaqMan Fast Universal PCR Master Mix (2?), No AmpErase UNG, instead of the TaqMa...
Instructions for setting up the computer system prior to instrument installation.
Provides a recommended workflow and timeline for the installation process.
Instructions for safely uncrating and positioning the instrument.
Procedure to verify that all required components have been received.
Steps to physically set up the instrument components.
Detailed steps for connecting and powering up the system.
Step-by-step guide to installing the SDS software.
Instructions for powering on the instrument after software installation.
Guides on running and interpreting the instrument function test.
Steps for preparing the calibration plate.
Guides on creating the necessary plate document in the software.
Detailed steps for executing the ROI calibration process.
Instructions for preparing the background calibration plate.
Guides on creating the plate document for background calibration.
Step-by-step guide for performing the background calibration.
Procedure for analyzing the data obtained from background calibration.
Specific steps for optical calibration on 7500/7500 Fast systems.
Guides on creating the plate document for optical calibration.
Detailed steps for executing the optical calibration process.
Procedure for analyzing the data obtained from optical calibration.
Steps to prepare for performing the pure dye calibration.
Guides on creating the plate document for pure dye calibration.
Instructions for preparing plates and running the calibration.
Procedure for analyzing the data from pure dye calibration runs.
Guides on verifying the spectral peaks for pure dyes.
Steps for preparing the RNase P verification plate.
Guides on creating the plate document for RNase P verification.
Instructions for running the instrument verification test.
Solutions for issues encountered during verification.
General| Type | Real-Time PCR System |
|---|---|
| Detection Method | Fluorescence |
| Weight | Not found in official specification |
| Temperature Uniformity | ±0.5°C |
| Dimensions (W x D x H) | Not found in official specification |
| Detection Channels | 4 |
| Sample Capacity | 96-well |
| Temperature Range | 4°C to 100°C |
| Temperature Accuracy | ±0.25°C |
| Ramp Rate (heating) | 3.5°C/sec |
| Ramp Rate (cooling) | 2.5°C/sec |
| Thermal Cycler Block | 96-well |
| Heating Rate | 3.5°C/sec |
| Cooling Rate | 2.5°C/sec |
| Power Requirements | 100-240 V, 50-60 Hz |
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