| 1. |
- Erni, W., et al.
(författare)
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Technical design report for the PANDA (AntiProton Annihilations at Darmstadt) Straw Tube Tracker
- 2013
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Ingår i: European Physical Journal A. Hadrons and Nuclei. - : Springer Science and Business Media LLC. - 1434-6001 .- 1434-601X. ; 49:2
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Tidskriftsartikel (refereegranskat)abstract
- This document describes the technical layout and the expected performance of the Straw Tube Tracker (STT), the main tracking detector of the PANDA target spectrometer. The STT encloses a Micro-Vertex-Detector (MVD) for the inner tracking and is followed in beam direction by a set of GEM stations. The tasks of the STT are the measurement of the particle momentum from the reconstructed trajectory and the measurement of the specific energy loss for a particle identification. Dedicated simulations with full analysis studies of certain proton-antiproton reactions, identified as being benchmark tests for the whole PANDA scientific program, have been performed to test the STT layout and performance. The results are presented, and the time lines to construct the STT are described.
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| 2. |
- Jemt, Anders, 1985-, et al.
(författare)
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Evaluation of methods for whole genome and transcriptome sequencing from nanograms of FFPE samples
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- The most widely used method for the preservation of clinical tissue specimens is formalin fixation and paraffin embedding (FFPE). Simultaneous analysis of RNA and DNA from samples preserved using this method have long proved problematic, primarily due to lack of material. Here, we describe an attempt to build a complete analysis package for RNA and DNA extracted from single tissue sections. The workflow includes quality control of the extracted material, library preparation and data analysis. We extract DNA with varying integrity from FFPE sections and subject them to whole genome sequencing using two library preparation methods, Illumina TruSeq Nano using the Illumina NeoPrep and Rubicon Genomics ThruPlex. We are able to obtain some usable data, albeit with high duplication rates, demonstrating both the possibilities and challenges of sequencing damaged DNA. Two different approaches to transcriptome sequencing are assessed, the TruSeq RNA Access library preparation kit from Illumina and the SMARTer Stranded Total RNA-Seq Kit - Pico Input from Clonetech. The sequence capture approach of the TruSeq kit is shown to be more robust to low integrity RNA compared to the SMARTer kit. However, the SMARTer kit needs much less starting material and is able to yield data about all transcripts, not just protein coding mRNA.
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| 3. |
- Barabash, Stas, et al.
(författare)
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Mars Orbiting Plasma Surveyor (MOPS)
- 2006
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Ingår i: Proceedings of the 6th IAA International Conference on Low-Cost Planetary Missions. ; , s. 227-232
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Mars Orbiting Plasma Surveyor (MOPS) is a microsatellite mission focused on studies of the near -Mars environment and the planet - solar wind interaction. The recent findings by the ESA Mars Express mission further highlighted the complexity of the processes taking place at the planet resulting from the solar wind interaction that strongly affect the planet's atmosphere. However, despite many previous Martian missions carrying different types of space plasma experiments, a comprehensive investigation including simultaneous measurements of particles, fields, and waves has never been performed. We consider a spinning spacecraft of a wet mass of 76.1 kg with a 9.7 kg payload, which can “hitchhike” on another platform until Mars orbit insertion, and then be released into a suitable orbit. The spacecraft design is based on the experience gained in very successful Swedish space plasma missions, Viking, Freja, Astrid-1, and Astrid-2. In the present mission design, the MOPS spacecraft is equipped with its own 1m high gain antenna for direct communication with the Earth. The payload includes a wave experiment with wire booms, magnetometer with a rigid boom, Langmuir probes, electron and ion energy spectrometers and an ion mass analyzer. An energetic neutral atom imager and an UV photometer may complete the core payload. One of the proposed scenarios is piggy - backing on the Russian Phobos - Grunt mission to be launched to Mars in 2011.
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| 4. |
- Borgström, Erik, et al.
(författare)
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Large Scale Library Generation for High Throughput Sequencing Authors and Affiliations
- 2011
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 6:4, s. e19119-
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Tidskriftsartikel (refereegranskat)abstract
- Background: Large efforts have recently been made to automatethe sample preparation protocols for massively parallel sequencing in order to match the increasing instrument throughput. Still, the size selection through agarose gel electrophoresis separation is a labor-intensive bottleneck of these protocols. Methodology/Principal Findings: In this study a method for automatic library preparation and size selection on a liquid handling robot is presented. The method utilizes selective precipitation of certain sizes of DNA molecules on to paramagnetic beads for cleanup and selection after standard enzymatic reactions. Conclusions/Significance: The method is used to generate libraries for de novo and re-sequencing on the Illumina HiSeq 2000 instrument with a throughput of 12 samples per instrument in approximately 4 hours. The resulting output data show quality scores and pass filter rates comparable to manually prepared samples. The sample size distribution can be adjusted for each application, and are suitable for all high throughput DNA processing protocols seeking to control size intervals.
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| 5. |
- Borgström, Erik, et al.
(författare)
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Phasing of single DNA molecules by massively parallel barcoding
- 2015
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Ingår i: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 6
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Tidskriftsartikel (refereegranskat)abstract
- High-throughput sequencing platforms mainly produce short-read data, resulting in a loss of phasing information for many of the genetic variants analysed. For certain applications, it is vital to know which variant alleles are connected to each individual DNA molecule. Here we demonstrate a method for massively parallel barcoding and phasing of single DNA molecules. First, a primer library with millions of uniquely barcoded beads is generated. When compartmentalized with single DNA molecules, the beads can be used to amplify and tag any target sequences of interest, enabling coupling of the biological information from multiple loci. We apply the assay to bacterial 16S sequencing and up to 94% of the hypothesized phasing events are shown to originate from single molecules. The method enables use of widely available short-read-sequencing platforms to study long single molecules within a complex sample, without losing phase information.
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| 6. |
- Fasterius, Erik, et al.
(författare)
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A novel RNA sequencing data analysis method for cell line authentication
- 2017
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Ingår i: PLOS ONE. - : PUBLIC LIBRARY SCIENCE. - 1932-6203. ; 12:2
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Tidskriftsartikel (refereegranskat)abstract
- We have developed a novel analysis method that can interrogate the authenticity of biological samples used for generation of transcriptome profiles in public data repositories. The method uses RNA sequencing information to reveal mutations in expressed transcripts and subsequently confirms the identity of analysed cells by comparison with publicly available cell-specific mutational profiles. Cell lines constitute key model systems widely used within cancer research, but their identity needs to be confirmed in order to minimise the influence of cell contaminations and genetic drift on the analysis. Using both public and novel data, we demonstrate the use of RNA-sequencing data analysis for cell line authentication by examining the validity of COLO205, DLD1, HCT15, HCT116, HKE3, HT29 and RKO colorectal cancer cell lines. We successfully authenticate the studied cell lines and validate previous reports indicating that DLD1 and HCT15 are synonymous. We also show that the analysed HKE3 cells harbour an unexpected KRAS-G13D mutation and confirm that this cell line is a genuine KRAS dosage mutant, rather than a true isogenic derivative of HCT116 expressing only the wild type KRAS. This authentication method could be used to revisit the numerous cell line based RNA sequencing experiments available in public data repositories, analyse new experiments where whole genome sequencing is not available, as well as facilitate comparisons of data from different experiments, platforms and laboratories.
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| 7. |
- Lindstrand, Anna, et al.
(författare)
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From cytogenetics to cytogenomics : whole-genome sequencing as a first-line test comprehensively captures the diverse spectrum of disease-causing genetic variation underlying intellectual disability
- 2019
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Ingår i: Genome Medicine. - : BMC. - 1756-994X. ; 11:1
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Tidskriftsartikel (refereegranskat)abstract
- BackgroundSince different types of genetic variants, from single nucleotide variants (SNVs) to large chromosomal rearrangements, underlie intellectual disability, we evaluated the use of whole-genome sequencing (WGS) rather than chromosomal microarray analysis (CMA) as a first-line genetic diagnostic test.MethodsWe analyzed three cohorts with short-read WGS: (i) a retrospective cohort with validated copy number variants (CNVs) (cohort 1, n=68), (ii) individuals referred for monogenic multi-gene panels (cohort 2, n=156), and (iii) 100 prospective, consecutive cases referred to our center for CMA (cohort 3). Bioinformatic tools developed include FindSV, SVDB, Rhocall, Rhoviz, and vcf2cytosure.ResultsFirst, we validated our structural variant (SV)-calling pipeline on cohort 1, consisting of three trisomies and 79 deletions and duplications with a median size of 850kb (min 500bp, max 155Mb). All variants were detected. Second, we utilized the same pipeline in cohort 2 and analyzed with monogenic WGS panels, increasing the diagnostic yield to 8%. Next, cohort 3 was analyzed by both CMA and WGS. The WGS data was processed for large (>10kb) SVs genome-wide and for exonic SVs and SNVs in a panel of 887 genes linked to intellectual disability as well as genes matched to patient-specific Human Phenotype Ontology (HPO) phenotypes. This yielded a total of 25 pathogenic variants (SNVs or SVs), of which 12 were detected by CMA as well. We also applied short tandem repeat (STR) expansion detection and discovered one pathologic expansion in ATXN7. Finally, a case of Prader-Willi syndrome with uniparental disomy (UPD) was validated in the WGS data.Important positional information was obtained in all cohorts. Remarkably, 7% of the analyzed cases harbored complex structural variants, as exemplified by a ring chromosome and two duplications found to be an insertional translocation and part of a cryptic unbalanced translocation, respectively.ConclusionThe overall diagnostic rate of 27% was more than doubled compared to clinical microarray (12%). Using WGS, we detected a wide range of SVs with high accuracy. Since the WGS data also allowed for analysis of SNVs, UPD, and STRs, it represents a powerful comprehensive genetic test in a clinical diagnostic laboratory setting.
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| 8. |
- Lundin, Carolina, et al.
(författare)
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Membrane topology of the Drosophila OR83b odorant receptor
- 2007
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Ingår i: FEBS Letters. - : Wiley. - 0014-5793 .- 1873-3468. ; 581:29, s. 5601-5604
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Tidskriftsartikel (refereegranskat)abstract
- By analogy to mammals, odorant receptors (ORs) in insects, such as Drosophila melanogaster, have long been thought to belong to the G-protein coupled receptor (GPCR) superfamily. However, recent work has cast doubt on this assumption and has tentatively suggested an inverted topology compared to the canonical N(out) - C(in) 7 transmembrane (TM) GPCR topology, at least for some Drosophila ORs. Here, we report a detailed topology mapping of the Drosophila OR83b receptor using engineered glycosylation sites as topology markers. Our results are inconsistent with a classical GPCR topology and show that OR83b has an intracellular N-terminus, an extracellular C-terminus, and 7TM helices.
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| 9. |
- Lundin, Daniel, et al.
(författare)
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Cross-field ion transport during high power impulse magnetron sputtering
- 2008
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Ingår i: Plasma sources science & technology. - : IOP Publishing. - 0963-0252 .- 1361-6595. ; 17:3
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Tidskriftsartikel (refereegranskat)abstract
- In this study, the effect on thin film growth due to an anomalous electron transport, found in high power impulse magnetron sputtering (HiPIMS), has been investigated for the case of a planar circular magnetron. An important consequence of this type of transport is that it affects the way ions are being transported in the plasma. It was found that a significant fraction of ions are transported radially outwards in the vicinity of the cathode, across the magnetic field lines, leading to increased deposition rates directly at the side of the cathode ( perpendicular to the target surface). Furthermore, this mass transport parallel to the target surface leads to that the fraction of sputtered material reaching a substrate placed directly in front of the target is substantially lower in HiPIMS compared with conventional direct current magnetron sputtering (dcMS). This would help to explain the lower deposition rates generally observed for HiPIMS compared with dcMS. Moreover, time-averaged mass spectrometry measurements of the energy distribution of the cross-field transported ions were carried out. The measured distributions show a direction-dependent high-energy tail, in agreement with predictions of the anomalous transport mechanism.
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| 10. |
- Lundin, Daniel, et al.
(författare)
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Transition between the discharge regimes of high power impulse magnetron sputtering and conventional direct current magnetron sputtering
- 2009
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Ingår i: Plasma sources science & technology. - : IOP Publishing. - 0963-0252 .- 1361-6595. ; 18:4
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Tidskriftsartikel (refereegranskat)abstract
- Current and voltage have been measured in a pulsed high power impulse magnetron sputtering (HiPIMS) system for discharge pulses longer than 100 mu s. Two different current regimes could clearly be distinguished during the pulses: (1) a high-current transient followed by (2) a plateau at lower currents. These results provide a link between the HiPIMS and the direct current magnetron sputtering (DCMS) discharge regimes. At high applied negative voltages the high-current transient had the characteristics of HiPIMS pulses, while at lower voltages the plateau values agreed with currents in DCMS using the same applied voltage. The current behavior was found to be strongly correlated with the chamber gas pressure, where increasing gas pressure resulted in increasing peak current and plateau current. Based on these experiments it is suggested here that the high-current transients cause a depletion of the working gas in the area in front of the target, and thereby a transition to a DCMS-like high-voltage, lower current regime.
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