Nature Methods Application Notes e-UPDATE: 9 December 2014

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9 December 2014  Application Notes bring you the latest information about innovative tools and technologies and their applications in the lab. We hope that you will find this service useful and informative and encourage you to sign up for future Updates to ensure that you never miss one!

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FEATURED APPLICATION NOTE QuantSeq 3′ mRNA sequencing for RNA quantification www.lexogen.com > QuantSeq provides an easy protocol to generate highly strand-specific next-generation sequencing (NGS) libraries close to the 3′ end of polyadenylated RNAs within 4.5 h. Only one fragment per transcript is generated, directly linking the number of reads mapping to a gene to its expression. QuantSeq reduces data analysis time and enables a higher level of multiplexing per run. QuantSeq is the RNA sample preparation method for accurate and affordable gene expression measurement. PDF

		Fast and Convenient 5-hydroxymethylcytosine Enrichment Workflow for Next-Generation Sequencing www.thermofisher.lt > 5-hydroxymethylcytosine (5-hmC) is an extensively studied DNA epigenetic modification. Here we present a novel tool which combined with next generation sequencing offers new ways of analyzing 5-hmC at the genomic level. We demonstrate that the Thermo Scientific™ EpiJET™ 5-hmC Enrichment Kit is highly specific for different DNA samples containing 5-hmC modifications. Our data shows that this tool can be used for locus-specifiand whole genome-specific analysis resulting in 5-hmC distribution patterns across different genomes. PDF

		Bioluminescent Nicotinamide Adenine Dinucleotide Detection Assays: Part II Choosing the Right Product for Your Application www.promega.com > The NAD(P)/NAD(P)H-Glo™ Assays are bioluminescent assays for the sensitive measurement of nicotinamide adenine dinucleotides. In this article we provide general guidelines for using these assays to measure enzyme activity in biochemical applications and for determining total or individual oxidized and reduced forms in cell-based applications. PDF

		Bioluminescent Nicotinamide Adenine Dinucleotide Detection Assays Part 1: Technology and Features www.promega.com > Here we introduce the NAD(P)/NAD(P)H-Glo™ Assays. These sensitive bioluminescent assays can be used to measure nicotinamide adenine dinucleotides in biochemical and cell-based applications. PDF

		Using PrimeScript™ Reverse Transcriptase for Quantification of shRNA Knockdown of a Low Abundance Target www.clontech.com > PrimeScript Reverse Transcriptase is a high efficiency enzyme that is ideal for preparing cDNA for the analysis of low abundance transcripts. In this experiment, Cdc14A mRNA, which is expressed at low levels, was quantified in mouse embryonic fibroblast (MEF) cells that had been transduced with either a control shRNA or a Cdc14A-specific shRNA. The PrimeScript RT Reagent Kit (Perfect Real Time) (Cat. #RR037A)and a reverse transcriptase from another supplier (Company I) were used to generate cDNA templates for measuring Cdc14A expression by real-time PCR using SYBR® Green detection. PDF

		Improved Target Selection Using xGen™ Lockdown™ Probes www.idtdna.com > Target enrichment via in-solution hybrid capture using xGen™ Lockdown™ Probes yields >5000-fold enrichment, excellent uniformity, and virtually no GC bias. Lockdown Probes are suitable for supplementing existing array-derived capture panels or creating entire custom standalone panels. PDF

		Chromatrap® 96: a new solid-state platform for high-throughput ChIP www.chromatrap.com > Chromatrap® 96 (C96) is a high-throughput analysis platform that profiles up to 96 transcription factors and epigenetic modifications simultaneously in less than 1 d. It enables sensitive, selective and reproducible target amplification with excellent signal-to-noise ratios, even from samples as small as 0.1 μg. Compatible with automated handling, C96 allows simultaneous investigation of parallel epigenetic landscapes, offering unprecedented assay flexibility and speed. PDF

 

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