Data readout from Proseek Multiplex panels
Please note that the data analysis steps below will be taken care of for you if you decide to send your samples to our dedicated, highly skilled Proseek Multiplex Analysis Service, who will provide you with a comprehensive NPX data/quality control report after your study is completed.
If more extensive data analysis services are required, our team of Olink biostatisticians can help you with customized statistical analysis – see our statistical packages.
If you intend to run our panels in your own lab, or simply want to know more details, please read on below!
Detection and sample analysis is performed by high-throughput real-time PCR analysis using the Fluidigm® BioMark™ HD System. This PCR platform is a truly high capacity system which enables simultaneous detection of 96 analytes in 96 samples creating 9,216 datapoints from a single run. The BioMark HD platform uses microfluidic distribution of sample and assays requiring only 7 nL reactions and takes less than 3 hours to complete. The BioMark HD platform consists of a real-time PCR instrument, 96×96 Dynamic Array™ IFC for multiplexing the samples and biomarkers, and an IFC loader to load the 96×96 Dynamic Array IFC. More information can be found on the Fluidigm website.
(Fluidigm, BioMark, and Dynamic Array are trademarks or registered trademarks of Fluidigm Corporation).
Data generation and quality control
For easier data analysis, Olink Proteomics have, together with MultiD, developed a GenEx software module for purchase that can be used for data generated with the Proseek Multiplex panels and the Fluidigm BioMark real-time PCR instrument. Your result file is exported to the GenEx software using the Olink Wizard which pre-processes your raw data for subsequent statistical analysis. Quality controls as well as normalization are performed automatically, providing you with Normalized Protein eXpression (NPX) values for further data analysis. Visualization tools are available, including scatter plot, box and whisker plot or bar graph, which allow you to rapidly identify major differences across samples.
The Olink Wizard add-on feature for GenEx is an easy-to-use data import and pre-processing tool. The Wizard will guide you through all steps of importing data, validating data quality and normalization of your Proseek Multiplex data, preparing it for statistical analysis with the GenEx software. You can download the Wizard Guide here.
Detector Setup files
To obtain the biomarker list, import the panel-specific .plt file directly into the Fluidigm RT PCR software. Make sure to load the Primer Plate according to figure 3 in the Proseek Multiplex User Manual. Please note that the detection setup.plt and setup.plt files should be saved to your computer first before imported into the Fluidigm RT PCR software. The .plt files can be downloaded below (right-click and “Save link as..” to your computer or device).
Proseek Default Sample Setup.plt
Proseek Cardiometabolic Detector Setup.plt
Proseek Cell Regulation Detector Setup.plt
Proseek CVD I Detector Setup.plt
Proseek CVD II Detector Setup.plt
Proseek CVD III Detector Setup.plt
Proseek Development Detector Setup.plt
Proseek Immune Response Detector Setup.plt
Proseek Immuno-Onc I Detector Setup.plt
Proseek Inflammation I Detector Setup.plt
Proseek Metabolism Detector Setup.plt
Proseek Neurology I Detector Setup.plt
Proseek Oncology I v2 Detector Setup.plt
Proseek Oncology II Detector Setup.plt
Proseek Organ Damage Detector Setup.plt
Data Pre-processing Excel Macro file
For data normalization without the Olink Wizard for GenEx, Olink Proteomics has developed a data pre-processing Excel Macro, free for download. It will perform data analysis pre-processing, including normalization and QC control steps, generating NPX and linear NPX values just as the GenEx software with the Olink Wizard. Please note that the data pre-processing Excel file can handle only one Proseek Multiplex run at a time and is therefore not suited for analysis of studies across multiple plates. If several runs are to be analyzed, the GenEx software with Olink Wizard is strongly recommended.
The macro can be downloaded below (right-click and “Save link as..” to your computer or device).
Download Excel Macro >>