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Showing papers by "Agilent Technologies published in 2007"


Journal ArticleDOI
TL;DR: This protocol explains how to use Cytoscape to analyze the results of mRNA expression profiling, and other functional genomics and proteomics experiments, in the context of an interaction network obtained for genes of interest.
Abstract: Cytoscape is a free software package for visualizing, modeling and analyzing molecular and genetic interaction networks. This protocol explains how to use Cytoscape to analyze the results of mRNA expression profiling, and other functional genomics and proteomics experiments, in the context of an interaction network obtained for genes of interest. Five major steps are described: (i) obtaining a gene or protein network, (ii) displaying the network using layout algorithms, (iii) integrating with gene expression and other functional attributes, (iv) identifying putative complexes and functional modules and (v) identifying enriched Gene Ontology annotations in the network. These steps provide a broad sample of the types of analyses performed by Cytoscape.

2,313 citations


Journal ArticleDOI
TL;DR: Using chromatin immunoprecipitation analysis, this paper showed that genes methylated in cancer cells are specifically packaged with nucleosomes containing histone H3 trimethylated on Lys27.
Abstract: Many genes associated with CpG islands undergo de novo methylation in cancer. Studies have suggested that the pattern of this modification may be partially determined by an instructive mechanism that recognizes specifically marked regions of the genome. Using chromatin immunoprecipitation analysis, here we show that genes methylated in cancer cells are specifically packaged with nucleosomes containing histone H3 trimethylated on Lys27. This chromatin mark is established on these unmethylated CpG island genes early in development and then maintained in differentiated cell types by the presence of an EZH2-containing Polycomb complex. In cancer cells, as opposed to normal cells, the presence of this complex brings about the recruitment of DNA methyl transferases, leading to de novo methylation. These results suggest that tumor-specific targeting of de novo methylation is pre-programmed by an established epigenetic system that normally has a role in marking embryonic genes for repression.

1,153 citations


Journal ArticleDOI
TL;DR: In this paper, an integrated analysis of high-density oligonucleotide array CGH and gene expression profiling data from 155 multiple myeloma samples identified a promiscuous array of abnormalities contributing to the dysregulation of NF-kappaB in approximately 20% of patients.

1,002 citations


Journal ArticleDOI
TL;DR: The implementation of this framework in a software application, termed DRIM (discovery of rank imbalanced motifs), which identifies sequence motifs in lists of ranked DNA sequences, is demonstrated, demonstrating that the statistical framework embodied in the DRIM software tool is highly effective for identifying regulatory sequence elements in a variety of applications.
Abstract: Computational methods for discovery of sequence elements that are enriched in a target set compared with a background set are fundamental in molecular biology research. One example is the discovery of transcription factor binding motifs that are inferred from ChIP–chip (chromatin immuno-precipitation on a microarray) measurements. Several major challenges in sequence motif discovery still require consideration: (i) the need for a principled approach to partitioning the data into target and background sets; (ii) the lack of rigorous models and of an exact p-value for measuring motif enrichment; (iii) the need for an appropriate framework for accounting for motif multiplicity; (iv) the tendency, in many of the existing methods, to report presumably significant motifs even when applied to randomly generated data. In this paper we present a statistical framework for discovering enriched sequence elements in ranked lists that resolves these four issues. We demonstrate the implementation of this framework in a software application, termed DRIM (discovery of rank imbalanced motifs), which identifies sequence motifs in lists of ranked DNA sequences. We applied DRIM to ChIP–chip and CpG methylation data and obtained the following results. (i) Identification of 50 novel putative transcription factor (TF) binding sites in yeast ChIP–chip data. The biological function of some of them was further investigated to gain new insights on transcription regulation networks in yeast. For example, our discoveries enable the elucidation of the network of the TF ARO80. Another finding concerns a systematic TF binding enhancement to sequences containing CA repeats. (ii) Discovery of novel motifs in human cancer CpG methylation data. Remarkably, most of these motifs are similar to DNA sequence elements bound by the Polycomb complex that promotes histone methylation. Our findings thus support a model in which histone methylation and CpG methylation are mechanistically linked. Overall, we demonstrate that the statistical framework embodied in the DRIM software tool is highly effective for identifying regulatory sequence elements in a variety of applications ranging from expression and ChIP–chip to CpG methylation data. DRIM is publicly available at http://bioinfo.cs.technion.ac.il/drim.

687 citations


Journal ArticleDOI
TL;DR: Electron transfer dissociation is an excellent method for localization of phosphorylation sites and should be an integral component of any strategy for comprehensive phosphorylated peptides analysis.
Abstract: Electron transfer dissociation (ETD) is a recently introduced mass spectrometric technique that provides a more comprehensive coverage of peptide sequences and posttranslational modifications. Here, we evaluated the use of ETD for a global phosphoproteome analysis. In all, we identified a total of 1,435 phosphorylation sites from human embryonic kidney 293T cells, of which 1,141 (≈80%) were not previously described. A detailed comparison of ETD and collision-induced dissociation (CID) modes showed that ETD identified 60% more phosphopeptides than CID, with an average of 40% more fragment ions that facilitated localization of phosphorylation sites. Although our data indicate that ETD is superior to CID for phosphorylation analysis, the two methods can be effectively combined in alternating ETD and CID modes for a more comprehensive analysis. Combining ETD and CID, from this single study, we were able to identify 80% of the known phosphorylation sites in >1,000 phosphorylated peptides analyzed. A hierarchical clustering of the identified phosphorylation sites allowed us to discover 15 phosphorylation motifs that have not been reported previously. Overall, ETD is an excellent method for localization of phosphorylation sites and should be an integral component of any strategy for comprehensive phosphorylation analysis.

532 citations


Journal ArticleDOI
TL;DR: It is shown that targeting oligonucleotides released from programmable microarrays can be used to capture and amplify ∼10,000 human exons in a single multiplex reaction, and it is anticipated that highly multiplexed methods for targeted amplification will enable the comprehensive resequencing ofhuman exons at a fraction of the cost of whole-genome resequenced.
Abstract: A new generation of technologies is poised to reduce DNA sequencing costs by several orders of magnitude. But our ability to fully leverage the power of these technologies is crippled by the absence of suitable 'front-end' methods for isolating complex subsets of a mammalian genome at a scale that matches the throughput at which these platforms will routinely operate. We show that targeting oligonucleotides released from programmable microarrays can be used to capture and amplify approximately 10,000 human exons in a single multiplex reaction. Additionally, we show integration of this protocol with ultra-high-throughput sequencing for targeted variation discovery. Although the multiplex capture reaction is highly specific, we found that nonuniform capture is a key issue that will need to be resolved by additional optimization. We anticipate that highly multiplexed methods for targeted amplification will enable the comprehensive resequencing of human exons at a fraction of the cost of whole-genome resequencing.

509 citations


Journal ArticleDOI
TL;DR: A specially designed cantilever tip is created that allows these interaction forces to be measured with good (sub-microsecond) temporal resolution and material properties to be determined and mapped in detail with nanoscale spatial resolution.
Abstract: Tapping-mode atomic force microscopy (AFM), in which the vibrating tip periodically approaches, interacts and retracts from the sample surface, is the most common AFM imaging method. The tip experiences attractive and repulsive forces that depend on the chemical and mechanical properties of the sample, yet conventional AFM tips are limited in their ability to resolve these time-varying forces. We have created a specially designed cantilever tip that allows these interaction forces to be measured with good (sub-microsecond) temporal resolution and material properties to be determined and mapped in detail with nanoscale spatial resolution. Mechanical measurements based on these force waveforms are provided at a rate of 4 kHz. The forces and contact areas encountered in these measurements are orders of magnitude smaller than conventional indentation and AFM-based indentation techniques that typically provide data rates around 1 Hz. We use this tool to quantify and map nanomechanical changes in a binary polymer blend in the vicinity of its glass transition. Phase changes and chemical compositional variations in materials on the nanoscale have been studied using various scanning force microscopy techniques. In these techniques, a force-sensing cantilever with a sharp tip is placed in continuous contact with the sample surface. Dynamical properties of the cantilever are adjusted depending on the material in contact with the tip. Examples of these techniques include ultrasonic force microscopy 1 , force modulation microscopy 2 , shear modulation force microscopy and lateral force microscopy 3,4 .T hese

469 citations


Journal ArticleDOI
21 Jun 2007-Nature
TL;DR: The results indicate that murine and human tumours experience common biological processes driven by orthologous genetic events in their malignant evolution.
Abstract: Highly rearranged and mutated cancer genomes present major challenges in the identification of pathogenetic events driving the neoplastic transformation process. Here we engineered lymphoma-prone mice with chromosomal instability to assess the usefulness of mouse models in cancer gene discovery and the extent of cross-species overlap in cancer-associated copy number aberrations. Along with targeted re-sequencing, our comparative oncogenomic studies identified FBXW7 and PTEN to be commonly deleted both in murine lymphomas and in human T-cell acute lymphoblastic leukaemia/lymphoma (T-ALL). The murine cancers acquire widespread recurrent amplifications and deletions targeting loci syntenic to those not only in human T-ALL but also in diverse human haematopoietic, mesenchymal and epithelial tumours. These results indicate that murine and human tumours experience common biological processes driven by orthologous genetic events in their malignant evolution. The highly concordant nature of genomic events encourages the use of genomically unstable murine cancer models in the discovery of biological driver events in the human oncogenome.

392 citations


Journal ArticleDOI
01 Jan 2007-RNA
TL;DR: A sensitive, accurate, and multiplexed microRNA (miRNA) profiling assay that is based on a highly efficient labeling method and novel microarray probe design that allows rapid design and validation of probes for simultaneous quantitative measurements of all human miRNA sequences in the public databases and to new mi RNA sequences as they are reported.
Abstract: We have developed a sensitive, accurate, and multiplexed microRNA (miRNA) profiling assay that is based on a highly efficient labeling method and novel microarray probe design. The probes provide both sequence and size discrimination, yielding in most cases highly specific detection of closely related mature miRNAs. Using a simple, single-vial experimental protocol, 120 ng of total RNA is directly labeled using Cy3 or Cy5, without fractionation or amplification, to produce precise and accurate measurements that span a linear dynamic range from 0.2 amol to 2 fmol of input miRNA. The results can provide quantitative estimates of the miRNA content for the tissues studied. The assay is also suitable for use with formalin-fixed paraffin-embedded clinical samples. Our method allows rapid design and validation of probes for simultaneous quantitative measurements of all human miRNA sequences in the public databases and to new miRNA sequences as they are reported.

321 citations


Journal ArticleDOI
TL;DR: The use of chromatin immunoprecipitation coupled with genome-wide promoter microarrays to query the occupancy of three ETS proteins in a human T-cell line found a highly divergent binding site that facilitates ETS1 and RUNX1 cooperative DNA binding.
Abstract: The conservation of in vitro DNA-binding properties within families of transcription factors presents a challenge for achieving in vivo specificity. To uncover the mechanisms regulating specificity within the ETS gene family, we have used chromatin immunoprecipitation coupled with genome-wide promoter microarrays to query the occupancy of three ETS proteins in a human T-cell line. Unexpectedly, redundant occupancy was frequently detected, while specific occupancy was less likely. Redundant binding correlated with housekeeping classes of genes, whereas specific binding examples represented more specialized genes. Bioinformatics approaches demonstrated that redundant binding correlated with consensus ETS-binding sequences near transcription start sites. In contrast, specific binding sites diverged dramatically from the consensus and were found further from transcription start sites. One route to specificity was found—a highly divergent binding site that facilitates ETS1 and RUNX1 cooperative DNA binding. The specific and redundant DNA-binding modes suggest two distinct roles for members of the ETS transcription factor family.

303 citations


Journal ArticleDOI
TL;DR: Two approaches provide significant new insights into both tissue-specific and general transcriptional targets in a crucial Shh-mediated patterning process.
Abstract: Sonic hedgehog (Shh) acts as a morphogen to mediate the specification of distinct cell identities in the ventral neural tube through a Gli-mediated (Gli1-3) transcriptional network. Identifying Gli targets in a systematic fashion is central to the understanding of the action of Shh. We examined this issue in differentiating neural progenitors in mouse. An epitope-tagged Gli-activator protein was used to directly isolate cis-regulatory sequences by chromatin immunoprecipitation (ChIP). ChIP products were then used to screen custom genomic tiling arrays of putative Hedgehog (Hh) targets predicted from transcriptional profiling studies, surveying 50-150 kb of non-transcribed sequence for each candidate. In addition to identifying expected Gli-target sites, the data predicted a number of unreported direct targets of Shh action. Transgenic analysis of binding regions in Nkx2.2, Nkx2.1 (Titf1) and Rab34 established these as direct Hh targets. These data also facilitated the generation of an algorithm that improved in silico predictions of Hh target genes. Together, these approaches provide significant new insights into both tissue-specific and general transcriptional targets in a crucial Shh-mediated patterning process.

Journal ArticleDOI
TL;DR: The results show that the phenotypic abnormalities of apparently balanced de novo CCRs are mainly due to cryptic deletions and that spermatogenesis is more prone to generate multiple chaotic chromosome imbalances and reciprocal translocations than oogenesis.
Abstract: Using array comparative genome hybridisation (CGH) 41 de novo reciprocal translocations and 18 de novo complex chromosome rearrangements (CCRs) were screened. All cases had been interpreted as "balanced" by conventional cytogenetics. In all, 27 cases of reciprocal translocations were detected in patients with an abnormal phenotype, and after array CGH analysis, 11 were found to be unbalanced. Thus 40% (11 of 27) of patients with a "chromosomal phenotype" and an apparently balanced translocation were in fact unbalanced, and 18% (5 of 27) of the reciprocal translocations were instead complex rearrangements with >3 breakpoints. Fourteen fetuses with de novo, apparently balanced translocations, all but two with normal ultrasound findings, were also analysed and all were found to be normal using array CGH. Thirteen CCRs were detected in patients with abnormal phenotypes, two in women who had experienced repeated spontaneous abortions and three in fetuses. Sixteen patients were found to have unbalanced mutations, with up to 4 deletions. These results suggest that genome-wide array CGH may be advisable in all carriers of "balanced" CCRs. The parental origin of the deletions was investigated in 5 reciprocal translocations and 11 CCRs; all were found to be paternal. Using customized platforms in seven cases of CCRs, the deletion breakpoints were narrowed down to regions of a few hundred base pairs in length. No susceptibility motifs were associated with the imbalances. These results show that the phenotypic abnormalities of apparently balanced de novo CCRs are mainly due to cryptic deletions and that spermatogenesis is more prone to generate multiple chaotic chromosome imbalances and reciprocal translocations than oogenesis.

Proceedings ArticleDOI
09 Jul 2007
TL;DR: This paper will present a tutorial on the control of AFMs, taking the reader on a walk around the control loop and discussing each of the individual technology components.
Abstract: The atomic force microscope (AFM) is one of the most versatile tools in nanotechnology. For control engineers this instrument is particularly interesting, since its ability to image the surface of a sample is entirely dependent upon the use of a feedback loop. This paper will present a tutorial on the control of AFMs. We take the reader on a walk around the control loop and discuss each of the individual technology components. The major imaging modes are described from a controls perspective and recent advances geared at increasing the performance of these microscopes are highlighted.

Journal ArticleDOI
TL;DR: Genes involved in differentiation and development were significantly over-represented and approximately half of the genes identified feature in the Online Mendelian Inheritance in Man database, suggesting that they could have important implications for phenotype and, thus, be useful for association studies of complex diseases.
Abstract: The discovery of copy number variation in healthy individuals is far from complete, and owing to the resolution of detection systems used, the majority of loci reported so far are relatively large ( approximately 65%>10 kb). Applying a two-stage high-resolution array comparative genomic hybridization approach to analyse 50 healthy Caucasian males from northern France, we discovered 2208 copy number variants (CNVs) detected by more than one consecutive probe. These clustered into 1469 CNV regions (CNVRs), of which 721 are thought to be novel. The majority of these are small (median size 4.4 kb) and most have common boundaries, with a coefficient of variation less than 0.1 for 83% of endpoints in those observed in multiple samples. Only 6% of the CNVRs analysed showed evidence of both copy number losses and gains at the same site. A further 6089 variants were detected by single probes: 48% of these were observed in more than one individual. In total, 2570 genes were seen to intersect variants: 1284 in novel loci. Genes involved in differentiation and development were significantly over-represented and approximately half of the genes identified feature in the Online Mendelian Inheritance in Man database. The biological importance of many genes affected, along with the well-conserved nature of the majority of the CNVs, suggests that they could have important implications for phenotype and, thus, be useful for association studies of complex diseases.

Journal ArticleDOI
TL;DR: In this paper, a single-mode quantum cascade laser source is presented, which is suitable for a variety of chemical sensing applications and can be used to perform absorption spectroscopy of fluids.
Abstract: We demonstrate a compact, single-mode quantum cascade laser source continuously tunable between 8.7 and 9.4μm. The source consists of an array of single-mode distributed feedback quantum cascade lasers with closely spaced emission wavelengths fabricated monolithically on a single chip and driven by a microelectronic controller. Our source is suitable for a variety of chemical sensing applications. Here, we use it to perform absorption spectroscopy of fluids.

Journal ArticleDOI
TL;DR: A bowtie plasmonic quantum cascade laser antenna that can confine coherent mid-infrared radiation well below the diffraction limit and efficiently suppresses the field enhancement at the outer ends of the structure, making it more suitable for spatially-resolved high-resolution chemical and biological imaging and spectroscopy.
Abstract: We report a bowtie plasmonic quantum cascade laser antenna that can confine coherent mid-infrared radiation well below the diffraction limit. The antenna is fabricated on the facet of a mid-infrared quantum cascade laser and consists of a pair of gold fan-like segments, whose narrow ends are separated by a nanometric gap. Compared with a nano-rod antenna composed of a pair of nano-rods, the bowtie antenna efficiently suppresses the field enhancement at the outer ends of the structure, making it more suitable for spatially-resolved high-resolution chemical and biological imaging and spectroscopy. The antenna near field is characterized by an apertureless near-field scanning optical microscope; field confinement as small as 130 nm is demonstrated at a wavelength of 7.0 mum.

Journal ArticleDOI
TL;DR: It is shown that the Wnt/β-catenin pathway also is essential for cardiac myogenesis to occur in ES cells, acting at a gastrulation-like stage, mediating mesoderm formation and patterning (two prerequisites for cardiacMyogenesis itself).
Abstract: Early steps for cardiac specification are problematic for the study of mammalian embryos, which has favored using pluripotent cells that recapitulate cardiac myogenesis. Furthermore, circuits governing cardiac specification have relevance to the application of ES cells and other cells for heart repair. In mouse teratocarcinoma cells, canonical Wnts that inhibit heart formation in avian or amphibian embryos and explants activate cardiogenesis, paradoxically. Here, we show that the Wnt/β-catenin pathway also is essential for cardiac myogenesis to occur in ES cells, acting at a gastrulation-like stage, mediating mesoderm formation and patterning (two prerequisites for cardiac myogenesis itself). Among genes associated temporally with this step was Sox17, encoding an endodermal HMG-box transcription factor. Using lentiviral vectors for RNA interference in differentiating ES cells, an essential role for Sox17 was proven in cardiac muscle cell formation. Sox17 short-hairpin RNA suppresses cardiac myogenesis selectively, acting subsequent to mesoderm formation yet before induction of Mesp1 and Mesp2, a pair of related basic helix–loop–helix transcription factors that together are indispensable for creating heart mesoderm. Sox17 short-hairpin RNA blocks cardiac myogenesis non-cell autonomously and impairs the induction of Hex, a homeodomain transcription factor that is known to be required for the production of endoderm-derived heart-inducing factors.

Journal ArticleDOI
TL;DR: The design principle and fabrication processes of the Agilent HPLC-Chip are summarized and the applications published so far are reviewed.
Abstract: It has been over 3 years since the first publication of the polymer microfluidic HPLC-Chip technology and more than 1 year since this technology became commercially available. Here, we summarize the design principle and fabrication processes of the Agilent HPLC-Chip and review the applications published so far.

Journal ArticleDOI
TL;DR: The joint analysis of array CGH and gene expression analysis highlights genes with concordant changes in expression and copy number that may be critical to lung cancer development and progression.

Journal ArticleDOI
TL;DR: Experimental results are presented that show that potential accuracy limitations introduced by the physical layer of the IEEE 802.11b wireless local area network do not preclude clock-synchronization accuracy of several hundred nanoseconds.
Abstract: IEEE 1588 is a new standard to synchronize independent clocks running on separate nodes of a distributed measurement and control system. It is intended for high-accuracy implementations on compact systems such as a single subnet. This paper examines potential accuracy limitations introduced by the physical layer of the IEEE 802.11b wireless local area network. Experimental results are presented that show that these limitations do not preclude clock-synchronization accuracy of several hundred nanoseconds.

Journal ArticleDOI
TL;DR: In this paper, a coherent multimode instability in quantum cascade laser (QCL) was observed, which is driven by the same fundamental mechanism of Rabi oscillations as the elusive Risken-Nummedal-Graham-Haken (RNGH) instability predicted decades ago for ring lasers.
Abstract: We report the observation of a coherent multimode instability in quantum cascade lasers (QCLs), which is driven by the same fundamental mechanism of Rabi oscillations as the elusive Risken-Nummedal-Graham-Haken (RNGH) instability predicted $40\phantom{\rule{0.3em}{0ex}}\text{years}$ ago for ring lasers. The threshold of the observed instability is significantly lower than in the original RNGH instability, which we attribute to saturable-absorption nonlinearity in the laser. Coherent effects, which cannot be reproduced by standard laser rate equations, can play therefore a key role in the multimode dynamics of QCLs, and in lasers with fast gain recovery in general.

Journal ArticleDOI
TL;DR: These studies demonstrate that a Deans switch can be an effective modulator provided that modulation ratios greater than approximately 2.5 are employed.
Abstract: A microfluidic Deans switch was used as a comprehensive two-dimensional gas chromatography (GC×GC) modulator. The simplicity and wide temperature range of the Deans switch make it a promising alternative to existing modulation techniques. However, the Deans switch is a low duty cycle modulator; that is, it samples only a small portion of the primary column effluent. Like all low duty cycle modulators, the Deans switch produces inconsistent transfer of components from the primary to the secondary column if the primary peaks are undersampled. Theoretical simulations and experimental studies show that the relative standard deviation (RSD) of the fraction of material transferred from the primary column to the secondary column is less than 1% if the modulation ratio is greater than 2.5. But the RSDs increase rapidly as the modulation ratio is decreased below 2.5. Deans switch GC×GC was validated by analyzing the aromatic content of gasoline. A fast analysis (<10 min) produced narrow primary peaks and a modulat...

Journal ArticleDOI
TL;DR: In this article, the authors investigated factors influencing the success of ERP implementations in multinational manufacturing companies in the Malaysian Free Trade Zone and found that enterprise-wide communication and a project management program are key factors for ERP implementation, while other factors such as top management support and teamwork are not as critical to the outcome.
Abstract: Enterprise resource planning (ERP) implementations in multinational manufacturing companies have experienced various degrees of success. This article investigates factors influencing the success of ERP implementations in multinational manufacturing companies in the Malaysian Free Trade Zone. The results indicate that enterprise-wide communication and a project management program are key factors influencing the success of ERP implementations, while other factors such as top management support as well as teamwork and composition are not as critical to the outcome. Organizational culture is a moderator of the relationships between enterprise-wide communication, a project management program, and the success of ERP implementations.

Journal ArticleDOI
TL;DR: A new combined doping control screening method for the analysis of anabolic steroids in human urine using liquid chromatography/electrospray ionization orthogonal acceleration time-of-flight mass spectrometry (LCoaTOFMS and GCoaTOF MS) has been developed in order to acquire accurate full scan MS data to be used to detect designer steroids.
Abstract: A new combined doping control screening method for the analysis of anabolic steroids in human urine using liquid chromatography/electrospray ionization orthogonal acceleration time-of-flight mass spectrometry (LCoaTOFMS) and gas chromatography/electron ionization orthogonal acceleration time-of-flight mass spectrometry (GCoaTOFMS) has been developed in order to acquire accurate full scan MS data to be used to detect designer steroids. The developed method allowed the detection of representative prohibited substances, in addition to steroids, at concentrations of 10 ng/mL for anabolic agents and metabolites, 30 ng/mL for corticosteroids, 500 ng/mL for stimulants and fl-blockers, 250ng/mL for diuretics, and 200ng/mL for narcotics. Sample preparation was based on liquid-liquid extraction of hydrolyzed human urine, and the final extract was analyzed as trimethylsilylated derivatives in GCoaTOFMS and underivatized in LCoaTOFMS in positive ion mode. The sensitivity, mass accuracy, advantages and limitations of the developed method are presented. Copyright (C) 2007 John Wiley & Sons, Ltd.

Patent
15 Jun 2007
TL;DR: In this article, an automated system for purification of a substance of interest is described, which generally comprises an instrument for moving fluids through the system, a reagent pack for storing fluids, and a purification cartridge.
Abstract: The present invention provides an automated system for purification of a substance of interest. The system generally comprises an instrument for moving fluids through the system, a reagent pack for storing fluids, and a purification cartridge. The cartridge comprises two filtration units for binding substances based on different physical properties. The cartridge also comprises rotary valves for control of movement of fluids on the cartridge. In preferred embodiments, the system is useful for purifying RNA from blood samples.

Journal ArticleDOI
TL;DR: High abundance ions corresponding to various degrees of neutral water losses, as well as furylium ion production, dominate the CID spectra, and that the sequence-informative b and y ions were rarely observed when Amadori-modified peptides were fragmented is suggested.
Abstract: Non-enzymatic glycation of peptides and proteins by D-glucose has important implications in the pathogenesis of diabetes mellitus, particularly in the context of development of diabetic complications. The fragmentation behavior of glycated peptides produced from reaction of D-glucose with lysine residues was investigated by electron transfer dissociation (ETD) and collision-induced dissociation (CID) tandem mass spectrometry. It was found that high abundance ions corresponding to various degrees of neutral water losses, as well as furylium ion production, dominate the CID spectra, and that the sequence-informative b and y ions were rarely observed when Amadori-modified peptides were fragmented. Contrary to what was observed under CID conditions, ions corresponding to neutral losses of water or furylium ion production were not observed in the ETD spectra. Instead, abundant and almost complete series of c- and z-type ions were observed regardless of whether the modification site was located in the middle of the sequence or close to the N-terminus, greatly facilitating the peptide sequencing. This study strongly suggests that ETD is a better technique for proteomic studies of non-enzymatically glycated peptides and proteins.

Journal ArticleDOI
TL;DR: Preparation of monolithic capillary columns for separations in the CEC mode using UV-initiated polymerization of the plain monolith followed by functionalization of its pore surface by photografting has been studied.
Abstract: Preparation of monolithic capillary columns for separations in the CEC mode using UV-initiated polymerization of the plain monolith followed by functionalization of its pore surface by photografting has been studied. The first step enabled the preparation of generic poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths with optimized porous properties, controlled by the percentages of porogens 1-decanol and cyclohexanol in the polymerization mixture, irradiation time, and UV light intensity. Ionizable monomers [2-(methacryloyloxy)ethyl]trimethylammonium chloride or 2-acryloamido-2-methyl-1-propanesulfonic acid were then photografted onto the monolithic matrix, allowing us to control the direction of the EOF in CEC. Different strategies were applied to control the grafting density and, thereby, the magnitude of the EOF. To control the hydrophobic properties, two approaches were tested: (i) cografting of a mixture of the ionizable and hydrophobic monomers and (ii) sequential grafting of the ionizable and hydrophobic monomers. Cografting resulted in similar retention but higher EOF. With sequential grafting, more than 50% increase in retention factors was obtained and a slight decrease in EOF was observed due to shielding of the ionizable moieties.

Proceedings ArticleDOI
09 Jul 2007
TL;DR: An overview of the approaches explored in improving the control of AFMs, including Hinfin, lscr1, and model-inverse based methods are given, and advantages and disadvantages are discussed.
Abstract: The atomic force microscope (AFM) is a powerful imaging and nanofabrication tool that allows the user to observe and manipulate samples at the atomic level. However, one limitation of current AFMs is the long time required to obtain a quality image of a sample. Several researchers have investigated this problem in recent years, and we give an overview of the approaches explored, including Hinfin, lscr1, and model-inverse based methods. We compare and discuss advantages and disadvantages of the various approaches, and we end with a summary of open questions to be addressed in improving the control of AFMs.

Journal ArticleDOI
TL;DR: An ICP–MS instrument fitted with an octopole reaction system (ORS) was used to directly measure the inorganic contents of several biofuel materials, showing it to be a fast and very sensitive technique for the elemental analysis of biofuels.
Abstract: An ICP-MS instrument fitted with an octopole reaction system (ORS) was used to directly measure the inorganic contents of several biofuel materials. Following sample preparation by simple dilution in kerosene, the biofuels were analysed directly. The ORS effectively removed matrix- and plasma-based spectral interferences to enable measurement of all important analytes, including sulfur, at levels below those possible by ICP-OES. A range of commonly produced biofuels was analysed, and spike recovery and long-term stability data was acquired. Suitably configured ICP-MS has been shown to be a fast and very sensitive technique for the elemental analysis of biofuels.

Patent
16 Feb 2007
TL;DR: A microfluidic assembly (1;57) has at least one flow path (17,27,29,35,38;82,85,95) coupled to the flow path.
Abstract: A microfluidic assembly (1;57) has at least one microfluidic flow path (17,27,29,35,38;82,85,95) and at least one inlet port (11,13,15;81,97,101) coupled to the flow path (17,27,29,35,38;82,85,95). The microfluidic assembly (1;57) has a first microfluidic device (7;63) that executes a microfluidic process and has a second microfluidic device (9;61). The microfluidic assembly (1;57) has an interface (5;65). The interface (5;65) couples the first microfluidic device (7;63) and the second microfluidic device (9;61).