Friday, January 13, 2012

Nature Methods Application Notes e-UPDATE: 13 January 2012

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13 JANUARY 2012 
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FEATURED APPLICATION NOTE
"The Simple Western™: a gel-free, blot-free, hands-free Western blotting reinvention
www.proteinsimple.com >
Western blotting is considered the gold standard for protein detection and characterization. Although improvements to individual aspects of Western methodologies have been developed in recent years, none has integrated the entire process onto a single platform. ProteinSimple™ has developed Simple Western™ assays for protein sizing and quantitative immunodetection as an alternative to traditional Western blot analysis. Assays are performed on Simon™, an instrument that integrates and automates all manual operations associated with Western blotting.
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Duolink-“In-cell Co-IP” for visualization of protein interactions in situ
www.olink.com >
Duolink® offers a user-friendly solution for studying protein-protein interactions. Duolink utilizes a pair of antibodies that are capable of quantitatively reporting even weak and transient protein-protein interactions in natively expressing cells either as countable, bright fluorescent spots for a standard fluorescence microscope or via chromogenic detection for brightfield microscopes. It offers all the benefits of traditional coimmunoprecipitation (Co-IP) and western blotting but with better quantitative precision and more information about cell-to-cell variability and target localization. It is also amenable to high throughput in 96 or 384 wells.
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Generation of cell lines using Epstein-Barr Virus (EBV) transformation of small volumes of cryo-preserved whole blood and the use of bench-top flow cytometry to achieve high and reproducible success rates
www.hpacultures.org.uk >
ECACC has developed a procedure for the generation of lymphoblastoid cell lines (LCLs) by the direct EBV transformation of small amounts (800µl) of cryo-preserved whole blood eliminating any requirement for prior separation of peripheral blood lymphocytes (PBLs). The ability to directly transform frozen whole blood avoids the cost of separating and storing PBLs while retaining the option to make a cell line at any time from all or a subset of the collection. Consistent transformation success rates of over 90% have been achieved using bench-top flow cytometry to track lymphoblastoid proliferation from source blood that has been cryo-preserved for more than four years.
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ReliaPrep™ Large Volume HT gDNA Isolation System: gDNA isolation from blood samples
www.promega.com >
The ReliaPrep™ Large Volume HT gDNA Isolation System enables automated purification of genomic DNA (gDNA) from up to 96 samples of whole blood at a time. The system dramatically increases laboratory throughput while conserving reagents by scaling the purification chemistry automatically to each sample's input volume, providing DNA that is ready for storage or for use in downstream applications without a need to resuspend DNA pellets.
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EndoLISA®: a novel and reliable method for endotoxin detection
www.hyglos.de >
A new test for the sensitive detection of endotoxin has been developed, based on a lipopolysaccharide-selective, precoated microplate and a factor C–based detection reagent and presented in a complete kit format. The selective capture of lipopolysaccharide (LPS) is achieved using a phage-derived receptor protein exhibiting high affinity and high specificity for the conserved core region of LPS. After binding of sample-LPS to the microplate as the first stage of the assay, the original sample matrix is washed off, thereby eliminating potentially interfering components. In the second stage of the assay, LPS is detected by factor C in a process whereby the principal receptor of the Limulus amoebocyte coagulation cascade reacts with a fluorescence substrate. The new endotoxin test EndoLISA has a detection range from 0.05 EU/ml up to 500 EU/ml.
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Expresso® Cloning and Expression Systems: Expressioneering™ Technology streamlines recombinant protein expression<
www.lucigen.com/ >
The Expresso® Cloning and Expression Systems use Expressioneering™ Technology to dramatically increase the speed and efficiency of target gene cloning and soluble protein expression in Escherichia coli. With Expressioneering, PCR products are cloned instantly and directionally into preprocessed expression vectors without sample cleanup or enzyme treatment. The Expresso Rhamnose System is a single-host system ideal for high throughput, allowing cloning in an effortless afternoon and recombinant protein expression the next day.
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Ribo-Zero Gold Kit: improved RNA-seq results after removal of cytoplasmic and mitochondrial ribosomal RNA
www.epibio.com >
Ribosomal RNA (rRNA) constitutes the majority (>98%) of total RNA preparations. To avoid wasting sequencing reads, it is necessary to remove this abundant RNA before preparing RNA libraries for deep sequencing. We previously developed Ribo-Zero™ technology for cytoplasmic rRNA removal from intact and degraded RNA. Here, we describe the Ribo-Zero Gold Kit: an improvement to the original Ribo-Zero method that allows removal of mitochondrial as well as cytoplasmic rRNA from total RNA preparations.
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