The MIQE guidelines have set a new standard for publishing qPCR results to ensure integrity in the scientific literature and increase experimental transparency. One of the key guidelines focuses on the proper design and experimental validation of primer assays used in qPCR. This tech report describes the design and wet-lab validation of PrimePCR assays and how they comply with the MIQE guidelines.
The T-helper cell (Th17) pathway has been implicated in several autoimmune diseases and in cancer. New magnetic bead–based assays have been developed for several biomarkers in this pathway. The performance characteristics of these assays are evaluated from data on intra- and inter-assay precision, standard curve recovery, cross reactivity, and linearity of dilution.
Hexapeptide libraries, such as Bio-Rad’s ProteoMiner™ protein enrichment technology, can significantly increase sensitivity of shotgun proteomics analyses of diverse complex samples. Underlying this increased sensitivity is the partial removal of high-abundance proteins leading to an increased representation of low-abundance proteins — a phenomenon described as dynamic range compression. Currently, hexapeptide libraries have been mostly used to increase proteomic detection in fluid samples (eg: saliva, plasma, etc.), but not in cellular lysates.
Highly Efficient Lipoparticle Capture and SPR Binding Kinetics of a Membrane Protein Using the ProteOn™ XPR36 Protein Interaction Array System
The genes coding for membrane proteins, such as G-protein coupled receptors (GPCR), ion channels, and other membrane-bound enzymes make up 25–30% of the human genome and membrane proteins are estimated to represent 30–40% of the top drug targets. There is growing research, industrial, and commercial interest in the study of membrane proteins in general, and more specifically in immobilizing membrane proteins to various biosensor surfaces.
A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data
This tech report shows how qPCR can lead to erroneous conclusions regarding differences in MCM7 gene expression between normal and tumor human breast cancer samples if the key steps set out in the MIQE guidelines are not followed.
Use of the PROTEAN® i12™ IEF System for In-Gel Peptide Fractionation Prior to LC-MS and Comparison with Off-Gel Fractionation
This tech report compares in-gel and off-gel separation of proteins in terms of overall speed and simplicity of the workflow, resolution of the separation, and depth of proteome coverage provided. Results found that as well as higher peptide yields, in-gel separations were better suited to take full advantage of downstream analysis methods.
PROTEAN® i12™ IEF System: Independent Voltage and Current Control Enables Optimization of First-Dimension IEF Conditions
This tech report demonstrates the advantages of independently controlled voltage and current parameters for each focusing tray in optimizing IEF conditions for a complex, insoluble membrane protein fraction.
This report demonstrates that the TC10 automated cell counter is a suitable alternative to the Coulter Counter and hemocytometer for the preparation of custom Bio-Plex assays.
This tech report describes an automated method of RNA extraction using the Aurum total RNA 96 kit and its use in a magnetic–bead based automated process. Results show that throughput can be easily increased with the Aurum total RNA 96 kit and that the automation protocols yield more reliable results, particularly when high sample volumes are used.
Not all cDNA synthesis kits have the same capacity to protect the integrity of RNA. An optimal level of RNase inhibitor is critical for powerful RNase inhibition and efficient reverse transcription. The iScript™ cDNA synthesis kit comes with a 5x iScript reaction mix and iScript reverse transcriptase, which is also blended with a potent RNase inhibitor.