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Invitrogen Lentiarray Human CRISPR Library, 96-Well Plate User Guide
USER GUIDE Invitrogen™ LentiArray™ Human CRISPR Library, 96-well Plate Catalog Numbers A31931, A31932, A31933, A31934, A31935, A31936, A31937, A31938, A31939, A31940, A31941, A31942, A31943, A31944, A31945, A31946, A31947, A31948, and A31949 Doc. Part No. 100044720 Pub. No. MAN0016075 Rev. B WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety Data Sheets (SDSs) are available from thermofisher.com/support. Product description Invitrogen™ LentiArray™ Human CRISPR libraries consist of pre-defined collection of gene families for functional genomics screening in an arrayed format. Each library targets a subset of human genes with up to 4 sequence-verified distinct lentiviral gRNA constructs per gene, pooled in a single well in a 96-well format. The gRNAs are based on the latest research on gRNA design. The gRNAs included in the LentiArray™ libraries are designed to knockout all known isoforms of the target genes and are selected for maximum knockout efficiency without sacrificing specificity. Characteristic Description Product Invitrogen™ LentiArray™ Human CRISPR Lentivirus Library (see Table 1, for details) Amount 4 aliquots of 50 µL/well per gene target (200 µL total per gene target) Viral titer • Libraries are delivered with a range of average titer between 2×107–2×108 TU/mL by puromycin antibiotic selection. • We recommend using 1×108 TU/mL for starting MOI calculations, see “MOI determination for screens“ on page 2 for additional guidance. Lentiviral map • gRNA expression is driven by a U6 promoter. • Includes puromycin resistance gene to allow selection of transduced cells. Plate layout • Refer to the associated PDF file for the plate map of the specific LentiArray™ Human CRISPR library and to the associated Excel files for gRNA target information. -
Cytoscan Assay Manual Workflow User
CytoScan™ Assay USER GUIDE Manual Workflow Catalog Numbers 901835, 901834, 901859, 901860 Publication Number 703038 Revision 7 For Research Use Only. Not for use in diagnostic procedures. The information in this guide is subject to change without notice. DISCLAIMER TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT. Important Licensing Information This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all applicable Limited Use Label Licenses. Corporate entity Life Technologies | Carlsbad, CA 92008 USA | Toll Free in USA 1 800 955 6288 TRADEMARKS All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Microsoft, and Excel are either registered trademarks or trademarks of Microsoft Corporation in the United States and/or other countries. QIAGEN, Gentra and Puregene are registered trademarks of QIAGEN. Titanium is a trademark of Clontech Laboratories, Inc. Tough-Spots is a registered trademark of Diversified Biotech. ©2011, 2014, 2017 Thermo Fisher Scientific Inc. All rights reserved. Contents CHAPTER 1 Before you begin. 6 Introduction . 6 Assay warnings and precautions . 7 Genomic DNA general requirements and recommendations . 9 Sources of human genomic DNA . 10 Genomic DNA extraction/purification methods . 10 RNase treatment . 10 Controls . 10 Data analysis . 11 CHAPTER 2 CytoScan™ Assay procedure . 12 Temperature definitions . 12 Section 1: Prepare gDNA plate . 13 Dilute stock gDNA to working concentration . -
RNA Isolation and Purification for Every Sample, RNA Type, and Application Isolate and Purify RNA with Confidence
RNA isolation and purification For every sample, RNA type, and application Isolate and purify RNA with confidence RNA isolation is a crucial step in your quest to understand gene expression levels. With all the solutions that Thermo Fisher Scientific has to offer, you can be confident that you’re getting started on the right foot. Go ahead and push the limits of your research. We’ll be there to support you with robust RNA purification kits, trusted RNA tools, and experienced technical support, all backed by nearly 30 years of leadership and innovation in RNA technologies. • Isolate from any sample type, for any application • Obtain high-purity, intact RNA • Achieve high yields, even from small sample quantities Contents Methods of RNA isolation 4 RNA and sample types 5 RNA applications 6 Organic RNA extraction 8 Spin column RNA extraction 10 Lysate-based RNA extraction 11 Automated RNA purification 12 Transcriptome purification 14 mRNA purification 15 MicroRNA and small RNA purification 16 Viral RNA purification 18 RNA from FFPE samples 20 RNA isolation technology guide 22 Avoiding RNA degradation 26 Tips for handling RNA 27 RNA quantitation 28 Gene expression solutions 30 Gene expression research considerations 31 Superior cDNA synthesis for any application 32 Complete kit with flexible priming options 33 Which instrument fits your needs? 34 Which thermal cycler or PCR instrument fits your needs? 35 RNA technical resources 36 Services and support 37 Ordering information 38 Methods of RNA isolation For every application, sample, and RNA type For over three decades, our scientists have developed and professional support. Our RNA isolation products innovative and robust RNA isolation products designed include organic reagents, columns, sample lysate, and to make your job as a scientist easier. -
Incoming Letter: QIAGEN N.V
WACHTELL, LIPTON , ROSEN & KATZ MARTIN LIPTON STEVEN A. COHEN 51 W E S T 52N D S T R E E T DAVID E. SHAPIRO SABASTIAN V. NILES HERBERT M. WACHTELL DEBORAH L. PAUL DAMIAN G. DIDDEN ALISON ZIESKE PREISS THEODORE N. MIRVIS DAVID C. KARP NEW YORK, N.Y. 1 0 0 1 9 - 6 1 5 0 IAN B OC Z K O TIJANA J. DVORNIC EDWARD D. HERLIHY RICHARD K. KIM MATTHEW M. GUEST JENNA E. LEVINE DANIEL A. NEFF JOSHUA R. CAMMAKER TELEPHONE: (212) 403 -1000 DAVID E. KAHAN RYAN A. McLEOD ANDREW R. BROWNSTEIN MARK GORDON DAVID K. LAM ANITHA REDDY MARC WOLINSKY JOSEPH D. LARSON FACSIMILE: (212) 403 -2000 BENJAMIN M. ROTH JOHN L. ROBINSON STEVEN A. ROSENBLUM JEANNEMARIE O’BRIEN JOSHUA A. FELTMAN JOHN R. SOBOLEWSKI JOHN F. SAVARESE WAYNE M. CARLIN GEORGE A. KATZ (1965 -1 9 8 9) ELAINE P. GOLIN STEVEN WINTER SCOTT K. CHARLES STEPHEN R. D iPRIMA JAMES H. FOGELSON (1967 - 1 9 91) EMI L A. KLEINHAUS EMILY D. JOHNSON JODI J. SCHWARTZ NICHOLAS G. DEMMO LEONARD M. ROSEN (1965 - 2 0 14 ) KARESSA L. CAIN JACOB A. KLING ADAM O. EMMERICH IGOR KIRMAN RONALD C. CHEN RAAJ S. NARAYAN RALPH M. LEVENE JONATHAN M. MOSES OF C O UN S EL GORDON S. MOODIE VIKTOR SAPEZHNIKOV RICHARD G. MASON T. EIKO STANGE DONGJU SONG MICHAEL J. SCHOBEL MARTIN J.E. ARMS ERIC S. ROBINSON DAVID M. SILK JOHN F. LYNCH BRADLEY R. WILSON ELINA TETELBAUM MICHAEL H. BYOWITZ PATRICIA A. ROBINSON* ROBIN PANOVKA WILLIAM SAVITT GRAHAM W. -
DCAT MEMBER COMPANY MEETING LOCATOR V. 4
DCAT MEMBER COMPANY MEETING LOCATOR v. 4 THE BENJAMIN Sancilio Pharmaceuticals Company, Inc. Sri Krishna Pharmaceuticals Ltd. Amino Chemicals Ltd. Zydus Pharmaceuticals (USA) Inc. Apogee Pharma, Inc. C2 PHARMA INTERCONTINENTAL BARCLAY Calyx Chemicals & Pharmaceuticals Ltd. AbbVie* ChemCon GmbH ACIC Pharmaceuticals Inc. Concord Biotech Limited Advitech SA Dipharma Francis Srl Amneal Pharmaceuticals LLC DSM Sinochem Pharmaceuticals ALP Pharm F.I.S. - Fabbrica Italiana Sintetici S.p.A. AMRI Jost Chemical Co. Asymchem Inc. Jubilant Pharma Capsugel, Now a Lonza Company* PharmSource, A GlobalData Company CBC AMERICAS Corp. PolyPeptide Group CellMark USA, LLC ROHNER Inc. Charioteer Pharmaceutical Co., Ltd., Zhejiang FIFTY NYC, A AFFINA HOTEL Chemical and Pharmaceutical Solutions Chiral Quest Corp. Chartwell Pharmaceuticals, LLC Croda, Inc. HOTEL 48LEX DFE Pharma AB BioTechnologies, Inc. DPL-US AiPing Pharmaceutical, Inc. EQ Esteve Almac Evonik Corporation Aptuit LLC FAREVA SA AZAD Fine Chemicals Ltd. Flavine North America, Inc. Cambridge Isotope Laboratories, Inc. Formosa Laboratories, Inc. CMC Biologics Grifols International S.A. Groupe Parima Hainan Poly Pharm. Co., Ltd. Navin Fluorine International Limited Harris Pharmaceutical Qualicaps, Inc. Helm AG RC2 Pharma Connect LLC Hetero USA, Inc. Recipharm Hikal, Ltd. Recro Gainesville LLC Interchem Corporation Reed-Lane, Inc. Inventia Healthcare PVT LTD Please note: Some DCAT member companies have requested not to be listed in the locator. (*) indicates member companies with Business Meeting Spaces in more than one hotel. INTERCONTINENTAL BARCLAY CONT'D PiSA BioPharm, Inc. SPI Pharma Inc. Johnson Matthey Tapemark Kingchem Life Science LLC Unither Pharmaceutical Legacy Pharmaceutical Packaging Uquifa S.A. Lonza AG* Neuland Laboratories Ltd. LOTTE NY PALACE Orion Group AbbVie* Par Pharmaceutical, Inc. -
Proteomics Tech Note 5802
proteomics tech note 5802 A Practical Approach to Proteomics Sean Taylor, Katrina Academia, Anthony Alburo, Aran Paulus, Kate Smith, and Considering all the possibilities, it is likely that any genome can Tanis Correa, Bio-Rad Laboratories, Inc. 2000 Alfred Nobel Drive, Hercules, CA potentially give rise to an infinite number of proteomes. Because 94547 USA proteins, not genes, are ultimately responsible for the phenotypic Since the completion of the human genome project, changes in cells and tissues, the mechanisms of disease, aging, sequencing technologies have continued to evolve, providing and environmental effects cannot be elucidated solely by tools for the rapid sequencing of most model organism studying the genome. The targets of drugs and chemicals are genomes. Associated genomic and transcriptomic data from proteins, and only through a survey of the proteome can the microarray and real-time PCR technologies have yielded associated mechanisms be understood. Most importantly, the a wealth of new information and deeper understanding of differential expression of mRNA (up or down) can capture at most biological systems. This genomic information has opened 40% of the variation of protein expression (Tian et al. 2004). up the field of proteomics, allowing the identification and The initial goal of most proteomics projects is to identify and comparison of differentially expressed proteins, from bacteria determine differential protein expression between samples. Once to humans. The accumulated data show that changes a list of differentially expressed proteins has been established, the in mRNA levels account for less than half of the relative subsequent step is to perform a detailed analysis of individual expression differences observed between associated proteins. -
Human Cytomegalovirus UL141 Protein Interacts with CELF5 and Affects Viral DNA Replication
MOLECULAR MEDICINE REPORTS 17: 4657-4664, 2018 Human cytomegalovirus UL141 protein interacts with CELF5 and affects viral DNA replication FEI ZOU1,2*, ZHI-TAO LU3*, SHUANG WANG1, SI WU1, YING-YING WU1 and ZHENG-RONG SUN1 1Department of BioBank, Affiliated Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004; 2Department of Pediatrics, First Hospital of Jilin University, Changchun, Jilin 130021; 3Department of Pediatrics, Zhangjiagang First People's Hospital, Zhangjiagang, Jiangsu 215600, P.R. China Received June 14, 2017; Accepted January 5, 2018 DOI: 10.3892/mmr.2018.8419 Abstract. Human cytomegalovirus (HCMV) infection is the regulation of HCMV genomic DNA synthesis, which is a key primary viral cause of congenital abnormalities and mental step during HCMV infection leading to neurological disease. retardation in newborns. The HCMV UL141-encoded glyco- protein has been previously revealed to inhibit the cell-surface Introduction expression of cluster of differentiation (CD)155, CD122, tumor necrosis factor-related apoptosis-inducing ligand death Human cytomegalovirus (HCMV) is a ubiquitous herpes virus (TRAIL)-receptor 1 (R1) and TRAIL-receptor 2 (R2), thus with infection rate of ~50-80% in females of reproductive age protecting virally-infected cells by allowing them to escape from different regions of the world, including Australia, Canada, natural killer cell-mediated cytotoxicity. The present study United States, Sweden, Finland, Spain, United Kingdom and investigated the interaction between HCMV UL141 and Ghana (1). Following an HCMV infection, the primary clinical human fetal brain cDNA to elucidate the possible effects of manifestations in healthy people are asymptomatic reces- UL141 on the nervous system. The findings of the current sive or latent infections. -
Life Sciences in Utah Read
EDCUTAH INDUSTRY PROFILE | FY 18-19 LIFE SCIENCES IN UTAH MAJOR UNIVERSITIES AND COLLEGES MAJOR EMPLOYERS ON THE COVER 1 Utah State University 4 Westminster College 6 Utah Valley University 9 Dixie State University 1 Paragon Medical Edwards Lifesciences Sapien 3 Heart Valve 2 Weber State University 5 Salt Lake 7 Brigham Young University 10 Rocky Vista University Community College 2 USU Research The Edwards Sapien 3 Transcatheter Heart Valve is changing 3 University of Utah 8 Southern Utah University Foundation the world of aortic stenosis therapy. In 2016 the Sapien 3 3 Fresenius USA received FDA approval for expanded use in the treatment of Manufacturing 4 Allergan intermediate-risk patients. The valve has the highest survival (formerly Actavis) rate ever reported in a TAVI clinical trial. 5 ARUP Laboratories LOGAN 6 Bard Access Systems 1 1 1 2 7 BioFire Diagnostics 8 Blackrock Microsystem 9 Biomerics 10 Clinical OGDEN Innovations 2 2 3 11 GE Healthcare 4 12 Great Basin 3 4 A 3 Scientific 4 SALT LAKE CITY 13 ICU Medical 5 A 6 14 Lineagen B 15 Myriad Genetic 4 Laboratories C 16 Nelson Laboratories 5 34 17 Otto Bock PROVO Healthcare 6 7 18 Pra Health Personalized Medicine, Medical Devices, Sciences 19 Recursion Pharmaceuticals 20 Sorenson World Class Research Genomics 21 Stryker Industry • In 2015, Utah’s post-performance tax incentives 22 Teva Pharma • Utah is proud to be home to companies like Merit facilitated nearly 25,000 new jobs and more than ceuticals Industries, (formerly Anesta, Medical Systems, BD (Becton Dickinson), Edwards $65M in new state revenue. -
Developing Biodefense Ivds Is Still a Priority John Conroy
Beyond Clinical Applications Developing biodefense IVDs is still a priority John Conroy In a scary world, nonclinical diagnostic testing spies a few challenges and many opportunities. s anthrax scares go, this one had that false alerts have become a fact of 1 Environmental and biowarfare. The as benign a beginning and ending life for lawmakers and their staffs. "We report noted that new technologies A as possible in today's terror- get a few of these a day," the spokesman have emerged since the 9/11 attacks to obsessed world. A New York City mu- told the Houston Chronicle. improve the detection, prevention, and sician who returned from Africa in As these stories attest, challenges and surveillance of bioterrorist attacks. February with unprocessed animal skins opportunities abound for IVD firms in a These technologies include rapid tests tested positive for anthrax. The 44-year- world where infectious diseases and for smallpox, anthrax, and salmonella, old man bought the skins, which con- bioterrorism agents are just a plane as well as improved vaccines and in- tained anthrax spores, to make tradi- flight, mail drop, cargo dock, or car ride formation-awareness networks to ex- tional African drums, authorities said. away. Market applications include tests pedite tracking the symptoms of chem- Complaining of flu-like symptoms, the for biodefense, avian flu, West Nile ical and biological events. musician collapsed while on tour with virus, food, animals and pets, water qual- his dance troupe in Pennsylvania, where ity, and genetically modified food. Test- • Food microbiology. In 2003, the glob- he was hospitalized. ing formats include ELISA, rapid later- al market for food microbiology tests Notable for their slightly more prosa- al-flow technology, and real-time was an estimated $864 million with an ic origins, similar stories of viral, toxic, polymerase chain reaction (PCR). -
Superscript® III Platinum® SYBR® Green
Page 4 Troubleshooting Guide Problem Possible Cause Solution ® ® ® No amplification product; cDNA synthesis temperature too Lower incubation temperature. SuperScript III Platinum SYBR Green One-Step qRT-PCR Kit Relative fluorescent high, low priming efficiency Cat. no. 11736-051 Size: 100 reactions signal ≤ background or RT or cDNA primer blocked by Raise incubation temperature. Redesign primer(s). Cat. no. 11736-059 Size: 500 reactions no-template control secondary structure RNA has been damaged/degraded Replace RNA if necessary. Store at -20°C RNase contamination Maintain aseptic conditions; add RNase inhibitor. Description Poor sensitivity Not enough template RNA Increase concentration of template RNA; use 10 ng–1 μg total RNA. The SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR kit a one-step quantitative RT-PCR kit for the detection and Product detected at RNA has been damaged/degraded Replace RNA if necessary. quantification of RNA using real-time detection instruments. This system combines SuperScript® III Reverse Transcriptase higher than expected RNase contamination Maintain aseptic conditions; add RNase inhibitor. ® ® cycle number (RT) and Platinum Taq DNA Polymerase in a single enzyme mix, with SYBR Green I fluorescent dye in a separate 2X RT inhibitors are present in RNA Remove inhibitors in the RNA preparation by an additional 70% reaction mix. Both cDNA synthesis and PCR are performed in a single tube using gene-specific primers and either total RNA ethanol wash. Inhibitors of RT include SDS, EDTA, guanidium or mRNA. Reagents are provided for 100 or 500 amplification reactions of 50 μl each. salts, formamide, sodium phosphate and spermidine. Inefficient cDNA synthesis Adjust cDNA synthesis temperature and/or primer design. -
Proteome Profiling of Exosomes Purified from a Small Amount Of
proteomes Article Proteome Profiling of Exosomes Purified from a Small Amount of Human Serum: The Problem of Co-Purified Serum Components Mateusz Smolarz 1, Monika Pietrowska 1, Natalia Matysiak 2, Łukasz Miela ´nczyk 2 and Piotr Widłak 1,* 1 Maria Skłodowska-Curie Institute—Oncology Center, Gliwice Branch, 44-101 Gliwice, Poland; [email protected] (M.S.); [email protected] (M.P.) 2 Department of Histology and Cell Pathology, School of Medicine with Division of Dentistry in Zabrze, Medical University of Silesia, 41-800 Zabrze, Poland; [email protected] (N.M.); [email protected] (Ł.M.) * Correspondence: [email protected]; Tel.: +48-32-2789672 Received: 26 March 2019; Accepted: 26 April 2019; Published: 28 April 2019 Abstract: Untargeted proteomics analysis of extracellular vesicles (EVs) isolated from human serum or plasma remains a technical challenge due to the contamination of these vesicles with lipoproteins and other abundant serum components. Here we aimed to test a simple method of EV isolation from a small amount of human serum (<1 mL) using the size-exclusion chromatography (SEC) standalone for the discovery of vesicle-specific proteins by the untargeted LC–MS/MS shotgun approach. We selected the SEC fraction containing vesicles with the size of about 100 nm and enriched with exosome markers CD63 and CD81 (but not CD9 and TSG101) and analyzed it in a parallel to the subsequent SEC fraction enriched in the lipoprotein vesicles. In general, there were 267 proteins identified by LC–MS/MS in exosome-containing fraction (after exclusion of immunoglobulins), yet 94 of them might be considered as serum proteins. -
Thermo Fisher Scientific Inc. 168 Third Avenue Waltham, Massachusetts 02451, U.S.A
Thermo Fisher Scientific Inc. 168 Third Avenue Waltham, Massachusetts 02451, U.S.A. THERMO FISHER SCIENTIFIC INC. 2007 EMPLOYEES' STOCK PURCHASE PLAN AS AMENDED AND RESTATED (THE "ESPP") Prospectus for the employees of certain European Economic Area ("EEA") subsidiaries of Thermo Fisher Scientific Inc., subject to the applicable legislation in each country Pursuant to articles L. 412-1 and L. 621-8 of the Code monétaire et financier and its General Regulation, in particular articles 211-1 to 216-1 thereof, the Autorité des marchés financiers ("AMF") has attached visa number 18-540 dated November 28, 2018, onto this prospectus. This prospectus was established by the issuer and incurs the responsibility of its signatories. The visa, pursuant to the provisions of Article L. 621-8-1-I of the Code monétaire et financier, was granted after the AMF verified that the document is complete and comprehensible, and that the information it contains is consistent. The visa represents neither the approval of the worthiness of the operation nor the authentication of the financial and accounting information presented. This prospectus will be made available in printed form to employees of the EEA subsidiaries of Thermo Fisher Scientific Inc. based in countries in which an offering under the ESPP is considered a public offering, subject to the applicable legislation in each country, at the respective head offices of their employers. In addition, this prospectus along with summary translations (as applicable) will be posted on Thermo Fisher Scientific Inc.’s intranet and free copies will be available to the employees upon request by contacting the human resources departments of their employers.