LightCycler® Software 4.1 – Features and Functionality
Read in this article:
- Improved Workflow
- Integrated Data
- User Management
- Enhanced Analysis Module Options
- Nucleic Acid Quantification
The LightCycler® Software 4.1 is an optimized version of the Software 4.05 for the use of in vitro diagnostic applications (restricted to the EU and countries where approved for this purpose) on the LightCycler® 2.0 Instrument (> Serial Number 1415000).
In combination with LightCycler® 2.0 Instrument (< Serial Number 1415000) or lower instrument versions, the software is for research and other non-regulated applications only.
LightCycler® Software 4.1 contains a number of features with relevance to in vitro diagnostic use:
- Traceable Database for in vitro diagnostic applications: deletion of experiment or manipulation of raw data is not possible.
- Audit Trail (documentation): report changes force user to enter change note/reason and username
- User Management is IVD conform
- Kit-specific Roche-defined Macros for IVD use:
- Macros are delivered with the specific IVD assays
- Contain run protocol, analysis and report
- Contain all settings for a specific IVD parameter kit.
- A wizard guides the user through the process
- Changes of Roche settings within the Macro leads to the following notification on the report: User Developed or Modified Test Method
- Macro Installer: easy installation of Roche Macros at customer site
LightCycler® Software 4.1 meets the demands of researchers by providing advanced data analysis modules, efficient data management, and effective data protection. This sophisticated software lets you automate your routine analysis, yet highly customize your most demanding analysis.
Functionalities and features of the LightCycler® Software 4.1 for research applications are fully retained and partially optimized compared to LightCycler® Software 4.05 and 4.0.
The modular structure of LightCycler® Software 4.1 makes PCR experiments and data analysis extremely easy to perform. You can now create experimental protocols quickly by using templates and macros. These allow you to store individual items, such as a PCR protocol, sample list, or a specific analysis from an old experiment as templates and then apply them to a new experiment. In addition, the settings for a complete experiment can be stored as a macro and can be directly applied to new experiments. A helpful software wizard guides through the process of applying the macros.
- Data generated with LightCycler® Software 4.1 are safely stored in a database.
- Enhanced data security allows no manipulation of raw data.
- A “Navigator” facilitates data base entries.
- Easy to use query possibility to find data.
The software offers a flexible user-management system that lets you create an account for each user that specifies log-in name, password, and level of access to the software. The software lets you assign one of three roles to the user, each with a different level of access
- Standard User
- Expert User
- Local Administrator
Each user level is allowed to perform different software operations, such as creating and modifying experimental protocols, using macros, and viewing folders and objects.
Enhanced Analysis Module Options
Increase the precision of your data by using the improved software algorithms. View all your results at a glance by using the “Call” option, which lets you compare samples with all sorts of controls including positive and negative run controls, and sample specific internal controls. Streamline your analysis by using one of the enhanced analysis modules:
The Qualitative Detection module detects the presence of a particular target in unknown samples, based on whether the target is amplified or not. However, the module does not quantify the amount of target DNA present. LightCycler®Software 4.1 lets you perform a simple Qualitative Detection experiment without controls, or add positive, negative, or internal controls for a more reliable Qualitative Detection experiment. The instrument displays the results of a Qualitative Detection experiment clearly, as shown below.
The “Combined” value is based on the “Target” result and the “Control” result. The “Target” result reports the observed amplification profile (amplification or no amplification) of each sample. The “Control” result shows the “Success” or “Failure” of all controls that are relevant to the particular sample. If any of those controls fail, the “Control” column will read “Failure” for the sample(s) affected.
Absolute and Relative Quantification
To analyze quantification data, LightCycler® Instruments measure fluorescent signals during the exponential increase (log-linear) phase of PCR and use the crossing-point (CP) of the sample (i.e., the cycle at which the fluorescence of a sample rises above background fluorescence), to determine the presence, concentration, or relative concentration of target DNA in unknown samples.
Calculate the concentration of your target DNA or RNA in unknown samples based on the concentration of standard samples or by comparing the ratio of two nucleic acid sequences.
Melting Curve Analysis
Determine sample melting temperatures and analyze melting profiles to obtain sequence information from the amplified PCR product. The LightCycler® Software 4.05 offers two types of Melting Curve Analysis: Genotyping Analysis and Tm Calling Analysis.
Nucleic Acid Quantification
Determine the concentration of nucleic acids without doing an amplification reaction.