Next-generation sequencing technologies
|
|
- Mervyn Tucker
- 5 years ago
- Views:
Transcription
1 Next-generation sequencing technologies
2 NGS applications
3 Illumina sequencing workflow
4 Overview Sequencing by ligation Short-read NGS Sequencing by synthesis Illumina NGS Single-molecule approach Long-read NGS Synthetic approach
5 General principles of short-read NGS Construct a library of fragments Generate clonal template populations Massively parallel DNA sequencing reactions Analyze data
6 General principles of short-read NGS Construct a library of fragments Generate clonal template populations Massively parallel DNA sequencing reactions Analyze data
7 Library preparation Prepares sample nucleic acids for sequencing Fragmentation Generates double-stranded DNA flanked by Illumina adapters Generates the same general template structure, but variables include Insert size Adapter type Index for multiplexing
8 Library preparation: Overview Purified genomic DNA Fragment DNA Repair Ends Fragments < 800bp Add an A to the 3 Ends Ligate Paired-end adapters Blunt end fragments with 5 phosphorylated ends Genomic DNA Library QC Library Amplified DNA with adapters PCR bp fragments Size-select on Gel
9 Library preparation: Fragmentation
10 Library preparation: Fragmentation The size of the target DNA fragments in the final library is a key parameter for NGS library construction. Optimal library size is impacted by 1. the process of cluster generation: Short products amplify more efficiently than longer products. Longer library inserts generate larger, more diffuse clusters than short inserts. 2. the sequencing application: For example, PE for exome sequencing since more than 80% of human exomes are under 200bp.
11 Library preparation: Fragmentation Three approaches are available to fragment nucleic acids: 1. Physical: Acoustic shearing and sonication, main method for genomic DNA 2. Enzymatic: Non-specific endonucleases cocktails or Transposase tagmentation, a greater number of artifactual indels compared with the physical method, reduced sampling handling and preparation time 3. Chemical: Heat and divalent metal cation, reserved for mrna
12 Library preparation: Repair Ends
13 Library preparation: A-tailing
14 Library preparation: A-tailing To facilitate ligation to sequencing adapter To prevent self-ligation between blunt ended template molecules (concatermers), or between adapters (adapter dimers) T P P A A P P A A P P A A P T P P T
15 Library preparation: Adapter ligation
16 Library preparation: Y-shaped adaptors
17 Library preparation: Y-shaped adapters Y-shaped adapters Non Y-shaped adapters
18 Library preparation: Size-select on Gel 600bp area excised 300bp area excised
19 Library preparation: PCR Selectively enrich DNA fragments with adapters on both ends Amplify the amount of DNA in the library
20 Library preparation: PCR
21 Library preparation: QC Library QC by Agilent Bioanalyzer: gives size confirmation and visualizes unwanted products Lower marker 15bp Upper marker 1500bp
22 General principles of short-read NGS Construct a library of fragments Generate clonal template populations Massively parallel DNA sequencing reactions Analyze data
23 Cluster amplification: Flow cells
24 Cluster amplification: Flow cells Adapter-ligated library elements hybridize to complementary oligonucleotides on the surface of a flow cell. Each attached library fragment acted as a seed and is amplified to generate a clonal cluster containing thousands of identical fragments. Ideally, clusters are of similar size and spaced well apart from each other to achieve accurate resolution during imaging. In reality, DNA clusters are randomly distributed across the flow cell with many clusters in close proximity to neighboring clusters, if the sample is overloaded, making it difficult to discern individual clusters from each others and reducing the amount of information generated during the run.
25 Cluster amplification: Patterned flow cells
26 Cluster amplification: Patterned flow cells Patterned flow cell technology provides even cluster spacing and uniform feature size to deliver extremely high cluster densities. Clusters can only form in the nanowells, allowing accurate resolution of clusters during imaging.
27 Cluster amplification
28 Cluster amplification
29 Cluster amplification: Hybridization and extension
30 Cluster amplification: Denaturation
31 Cluster amplification: Anchor the template to the surface
32 Cluster amplification: Bridge amplification
33 Cluster amplification: Bridge amplification
34 Cluster amplification: Denaturation
35 Cluster amplification: Bridge amplification
36 Cluster amplification: Bridge amplification
37 Cluster amplification: P5 Linearization P7 P5
38 Cluster amplification: P5 Linearization
39 Cluster amplification: Blocking
40 Cluster amplification: Read1 sequencing
41 General principles of short-read NGS Construct a library of fragments Generate clonal template populations Massively parallel DNA sequencing reactions Analyze data
42 Sequencing by synthesis
43 Sequencing by synthesis
44 Single read, paired-end and read lengths Program the system to sequence a specific number of bases ( bases) Sequence the strands from both directions to achieve a total of e.g. 600 bases (2 300 bases)
45 Paired-end sequencing Longer read lengths improve 1) the overall length of contiguous sequence that can be assembled, and 2) the certainty of short read alignments. Several next-generation sequencers have offered increases in read length over time. Another improvement has resulted from paired-end sequencing, producing sequence data from both ends of each library fragment. Read pairs can be obtained by one of two mechanisms: 1) paired ends or 2) mate pairs.
46 Paired-end sequencing
47 Paired-end sequencing
48 Paired-end sequencing
49 Paired-end sequencing: P7 linearization
50 Paired-end sequencing
51 Paired-end sequencing Fragment length Advantage (a) paired-end (b) mate-pair < 1000 bp > 1000 bp Higher accuracy of alignments than a single-end read of the same length Providing a scaffold for de novo sequencing by long-range order and orientation
52 Illumina: Summary
53 Illumina platforms: Benchtop sequencers
54 Illumina platforms: Production-scale sequencers
55 Choosing a library type Single read library Unidirectional sequencing Compatible with only single-read flow cells Applications: ChIP-seq, mrna-seq for quantification, low-coverage resequencing
56 Choosing a library type Paired end library Uni or Bidirectional sequencing Compatible with both single-read and paired-end flow cells Applications: the most common library type, de novo assembly, structural variants detection, high-coverage resequencing
57 Choosing a library type Indexed libraries Uni or bidirectional sequencing Allows multiple libraries per lane Single-indexed libraries: adds up to 48 unique 6-base index 1 (i7) se quences to generate up to 48 uniquely tagged libraries. Dual-indexed libraries: adds up to 24 unique 8-base index 1 (i7) sequences and up to 16 unique 8-base index 2 (i5) sequences to generate up to 384 uniquely tagged libraries.
58 Single-indexed sequencing The single-indexed sequencing workflow applies to all Illumina sequencing platforms.
59 Dual-indexed sequencing on a pairedend flow cell Dual-indexed sequencing includes 2 index reads.
60 Dual-indexed adapters
61 Dual-indexed sequencing: Workflow A 7 dark-cycles
62 Dual-indexed sequencing: Workflow A
63 Dual-indexed sequencing: Workflow B
64 Reads and coverage The number of reads for a specific region is denoted depth or coverage
Lecture 7. Next-generation sequencing technologies
Lecture 7 Next-generation sequencing technologies Next-generation sequencing technologies General principles of short-read NGS Construct a library of fragments Generate clonal template populations Massively
More informationNext-generation sequencing technologies
Next-generation sequencing technologies Illumina: Summary https://www.youtube.com/watch?v=fcd6b5hraz8 Illumina platforms: Benchtop sequencers https://www.illumina.com/systems/sequencing-platforms.html
More informationHuman Genome Sequencing Over the Decades The capacity to sequence all 3.2 billion bases of the human genome (at 30X coverage) has increased
Human Genome Sequencing Over the Decades The capacity to sequence all 3.2 billion bases of the human genome (at 30X coverage) has increased exponentially since the 1990s. In 2005, with the introduction
More informationDeep Sequencing technologies
Deep Sequencing technologies Gabriela Salinas 30 October 2017 Transcriptome and Genome Analysis Laboratory http://www.uni-bc.gwdg.de/index.php?id=709 Microarray and Deep-Sequencing Core Facility University
More informationLibrary construction for nextgeneration sequencing: Overviews. and challenges
Library construction for nextgeneration sequencing: Overviews and challenges During this time, as sequencing technologies have improved and evolved, so too have methods for preparing nucleic acids for
More informationHigh Throughput Sequencing the Multi-Tool of Life Sciences. Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center
High Throughput Sequencing the Multi-Tool of Life Sciences Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center Complementary Approaches Illumina Still-imaging of clusters (~1000
More informationWet-lab Considerations for Illumina data analysis
Wet-lab Considerations for Illumina data analysis Based on a presentation by Henriette O Geen Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center Complementary Approaches Illumina
More informationConsiderations for Illumina library preparation. Henriette O Geen June 20, 2014 UCD Genome Center
Considerations for Illumina library preparation Henriette O Geen June 20, 2014 UCD Genome Center Diversity of applications De novo genome Sequencing ranscriptome Expression Splice Isoform bundance Genotyping
More informationHigh Throughput Sequencing the Multi-Tool of Life Sciences. Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center
High Throughput Sequencing the Multi-Tool of Life Sciences Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center DNA Technologies & Expression Analysis Cores HT Sequencing (Illumina
More informationIntegrated NGS Sample Preparation Solutions for Limiting Amounts of RNA and DNA. March 2, Steven R. Kain, Ph.D. ABRF 2013
Integrated NGS Sample Preparation Solutions for Limiting Amounts of RNA and DNA March 2, 2013 Steven R. Kain, Ph.D. ABRF 2013 NuGEN s Core Technologies Selective Sequence Priming Nucleic Acid Amplification
More informationGet to Know Your DNA. Every Single Fragment.
HaloPlex HS NGS Target Enrichment System Get to Know Your DNA. Every Single Fragment. High sensitivity detection of rare variants using molecular barcodes How Does Molecular Barcoding Work? HaloPlex HS
More informationGenome Resequencing. Rearrangements. SNPs, Indels CNVs. De novo genome Sequencing. Metagenomics. Exome Sequencing. RNA-seq Gene Expression
Genome Resequencing De novo genome Sequencing SNPs, Indels CNVs Rearrangements Metagenomics RNA-seq Gene Expression Splice Isoform Abundance High Throughput Short Read Sequencing: Illumina Exome Sequencing
More informationIncorporating Molecular ID Technology. Accel-NGS 2S MID Indexing Kits
Incorporating Molecular ID Technology Accel-NGS 2S MID Indexing Kits Molecular Identifiers (MIDs) MIDs are indices used to label unique library molecules MIDs can assess duplicate molecules in sequencing
More informationWelcome to the NGS webinar series
Welcome to the NGS webinar series Webinar 1 NGS: Introduction to technology, and applications NGS Technology Webinar 2 Targeted NGS for Cancer Research NGS in cancer Webinar 3 NGS: Data analysis for genetic
More informationReading Lecture 8: Lecture 9: Lecture 8. DNA Libraries. Definition Types Construction
Lecture 8 Reading Lecture 8: 96-110 Lecture 9: 111-120 DNA Libraries Definition Types Construction 142 DNA Libraries A DNA library is a collection of clones of genomic fragments or cdnas from a certain
More informationNextGen Sequencing Technologies Sequencing overview
Outline Conventional NextGen High-throughput sequencing (Next-Gen sequencing) technologies. Illumina sequencing in detail. Quality control. Sequence coverage. Multiplexing. FASTQ files. Shendure and Ji
More informationDNA concentration and purity were initially measured by NanoDrop 2000 and verified on Qubit 2.0 Fluorometer.
DNA Preparation and QC Extraction DNA was extracted from whole blood or flash frozen post-mortem tissue using a DNA mini kit (QIAmp #51104 and QIAmp#51404, respectively) following the manufacturer s recommendations.
More informationThe New Genome Analyzer IIx Delivering more data, faster, and easier than ever before. Jeremy Preston, PhD Marketing Manager, Sequencing
The New Genome Analyzer IIx Delivering more data, faster, and easier than ever before Jeremy Preston, PhD Marketing Manager, Sequencing Illumina Genome Analyzer: a Paradigm Shift 2000x gain in efficiency
More informationNB536: Bioinformatics
NB536: Bioinformatics Instructor Prof. Jong Kyoung Kim Department of New Biology Office: E4-613 E-mail: jkkim@dgist.ac.kr Homepage: https://scg.dgist.ac.kr Course website https://scg.dgist.ac.kr/index.php/courses
More informationMatthew Tinning Australian Genome Research Facility. July 2012
Next-Generation Sequencing: an overview of technologies and applications Matthew Tinning Australian Genome Research Facility July 2012 History of Sequencing Where have we been? 1869 Discovery of DNA 1909
More informationFactors affecting PCR
Lec. 11 Dr. Ahmed K. Ali Factors affecting PCR The sequences of the primers are critical to the success of the experiment, as are the precise temperatures used in the heating and cooling stages of the
More informationRIPTIDE HIGH THROUGHPUT RAPID LIBRARY PREP (HT-RLP)
Application Note: RIPTIDE HIGH THROUGHPUT RAPID LIBRARY PREP (HT-RLP) Introduction: Innovations in DNA sequencing during the 21st century have revolutionized our ability to obtain nucleotide information
More informationDNA. bioinformatics. genomics. personalized. variation NGS. trio. custom. assembly gene. tumor-normal. de novo. structural variation indel.
DNA Sequencing T TM variation DNA amplicon mendelian trio genomics NGS bioinformatics tumor-normal custom SNP resequencing target validation de novo prediction personalized comparative genomics exome private
More informationNext Generation Sequencing. Jeroen Van Houdt - Leuven 13/10/2017
Next Generation Sequencing Jeroen Van Houdt - Leuven 13/10/2017 Landmarks in DNA sequencing 1953 Discovery of DNA double helix structure 1977 A Maxam and W Gilbert "DNA seq by chemical degradation" F Sanger"DNA
More informationIllumina Sequencing Overview
Illumina Sequencing Overview Part # 15045845_Rev.C 2013 Illumina, Inc. All rights reserved. Illumina, IlluminaDx, BaseSpace, BeadArray, BeadXpress, cbot, CSPro, DASL, DesignStudio, Eco, GAIIx, Genetic
More informationIntroductory Next Gen Workshop
Introductory Next Gen Workshop http://www.illumina.ucr.edu/ http://www.genomics.ucr.edu/ Workshop Objectives Workshop aimed at those who are new to Illumina sequencing and will provide: - a basic overview
More informationSupplementary Information for:
Supplementary Information for: A streamlined and high-throughput targeting approach for human germline and cancer genomes using Oligonucleotide-Selective Sequencing Samuel Myllykangas 1, Jason D. Buenrostro
More informationUnique, dual-matched adapters mitigate index hopping between NGS samples. Kristina Giorda, PhD
Unique, dual-matched adapters mitigate index hopping between NGS samples Kristina Giorda, PhD 1 Outline NGS workflow and cross-talk Sources of sample cross-talk and mitigation strategies Adapter recommendations
More informationNext Gen Sequencing. Expansion of sequencing technology. Contents
Next Gen Sequencing Contents 1 Expansion of sequencing technology 2 The Next Generation of Sequencing: High-Throughput Technologies 3 High Throughput Sequencing Applied to Genome Sequencing (TEDed CC BY-NC-ND
More informationRecitation CHAPTER 9 DNA Technologies
Recitation CHAPTER 9 DNA Technologies DNA Cloning: General Scheme A cloning vector and eukaryotic chromosomes are separately cleaved with the same restriction endonuclease. (A single chromosome is shown
More informationIncreased transcription detection with the NEBNext Single Cell/Low Input RNA Library Prep Kit
be INSPIRED drive DISCOVERY stay GENUINE TECHNICAL NOTE Increased transcription detection with the NEBNext Single Cell/Low Input RNA Library Prep Kit Highly sensitive, robust generation of high quality
More informationFunctional Genomics Research Stream. Research Meetings: November 2 & 3, 2009 Next Generation Sequencing
Functional Genomics Research Stream Research Meetings: November 2 & 3, 2009 Next Generation Sequencing Current Issues Research Meetings: Meet with me this Thursday or Friday. (bring laboratory notebook
More informationTECH NOTE Ligation-Free ChIP-Seq Library Preparation
TECH NOTE Ligation-Free ChIP-Seq Library Preparation The DNA SMART ChIP-Seq Kit Ligation-free template switching technology: Minimize sample handling in a single-tube workflow >> Simplified protocol with
More informationContact us for more information and a quotation
GenePool Information Sheet #1 Installed Sequencing Technologies in the GenePool The GenePool offers sequencing service on three platforms: Sanger (dideoxy) sequencing on ABI 3730 instruments Illumina SOLEXA
More informationNGS Sample QC with the Agilent 2200 TapeStation. Rainer Nitsche Application Engineer Agilent Technologies, Inc.
NGS Sample QC with the Agilent 2200 TapeStation Rainer Nitsche Application Engineer Agilent Technologies, Inc. The Agilent 2100 Bioanalyzer First commercially available Lab-on-a-Chip product Introduced
More informationTranscriptomics analysis with RNA seq: an overview Frederik Coppens
Transcriptomics analysis with RNA seq: an overview Frederik Coppens Platforms Applications Analysis Quantification RNA content Platforms Platforms Short (few hundred bases) Long reads (multiple kilobases)
More informationChapter 7. DNA Microarrays
Bioinformatics III Structural Bioinformatics and Genome Analysis Chapter 7. DNA Microarrays 7.9 Next Generation Sequencing 454 Sequencing Solexa Illumina Solid TM System Sequencing Process of determining
More informationOverview of Next Generation Sequencing technologies. Céline Keime
Overview of Next Generation Sequencing technologies Céline Keime keime@igbmc.fr Next Generation Sequencing < Second generation sequencing < General principle < Sequencing by synthesis - Illumina < Sequencing
More informationLab methods: Exome / Genome. Ewart de Bruijn
Lab methods: Exome / Genome 27 06 2013 Ewart de Bruijn Library prep is only a small part of the complete DNA analysis workflow DNA isolation library prep enrichment flowchip prep sequencing bioinformatics
More informationImplementation of Automated Sample Quality Control in Whole Exome Sequencing
Journal of Life Sciences 11 (2017) 261-268 doi: 10.17265/1934-7391/2017.06.001 D DAVID PUBLISHING Implementation of Automated Sample Quality Control in Whole Exome Sequencing Elisa Viering 1, Jana Molitor
More informationyou can see that if if you look into the you know the capability kilobases per day, per machine kind of calculation if you do.
Functional Genomics Professor S Ganesh Department of Biological Sciences & Bioengineering Indian Institute of Technology Kanpur Lecture No 11 DNA Sequencing Methods Part 2 So welcome back to this course
More informationThe Expanded Illumina Sequencing Portfolio New Sample Prep Solutions and Workflow
The Expanded Illumina Sequencing Portfolio New Sample Prep Solutions and Workflow Marcus Hausch, Ph.D. 2010 Illumina, Inc. All rights reserved. Illumina, illuminadx, Solexa, Making Sense Out of Life, Oligator,
More informationNext-Generation Sequencing. Technologies
Next-Generation Next-Generation Sequencing Technologies Sequencing Technologies Nicholas E. Navin, Ph.D. MD Anderson Cancer Center Dept. Genetics Dept. Bioinformatics Introduction to Bioinformatics GS011062
More informationIllumina s Suite of Targeted Resequencing Solutions
Illumina s Suite of Targeted Resequencing Solutions Colin Baron Sr. Product Manager Sequencing Applications 2011 Illumina, Inc. All rights reserved. Illumina, illuminadx, Solexa, Making Sense Out of Life,
More informationSureSelect Target Enrichment for the Ion Proton TM Next Generation Sequencing System
SureSelect Target Enrichment for the Ion Proton TM Next Generation Sequencing System Demonstrated performance you can count on Christina Chiu Product Manager, SureSelect Kyeong Jeong Ph.D. R&D Scientist
More informationG E N OM I C S S E RV I C ES
GENOMICS SERVICES ABOUT T H E N E W YOR K G E NOM E C E N T E R NYGC is an independent non-profit implementing advanced genomic research to improve diagnosis and treatment of serious diseases. Through
More informationEPIGENTEK. EpiNext DNA Library Preparation Kit (Illumina) Base Catalog # P-1051 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiNext DNA Library Preparation Kit (Illumina) Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE Uses: The EpiNext DNA Library Preparation Kit (Illumina) is suitable for preparing a DNA library
More informationsolid S Y S T E M s e q u e n c i n g See the Difference Discover the Quality Genome
solid S Y S T E M s e q u e n c i n g See the Difference Discover the Quality Genome See the Difference With a commitment to your peace of mind, Life Technologies provides a portfolio of robust and scalable
More informationHigh Throughput Sequencing the Multi-Tool of Life Sciences. Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center
High Throughput Sequencing the Multi-Tool of Life Sciences Lutz Froenicke DNA Technologies and Expression Analysis Cores UCD Genome Center DNA Technologies & Expression Analysis Cores HT Sequencing (Illumina
More informationab High Sensitivity DNA Library Preparation Kit (For Illumina )
ab185905 High Sensitivity DNA Library Preparation Kit (For Illumina ) Instructions for Use For the preparation of a DNA library using sub-nanogram amounts of DNA input for next generation sequencing applications
More informationab High Sensitivity DNA Library Preparation Kit (For Illumina )
ab185905 High Sensitivity DNA Library Preparation Kit (For Illumina ) Instructions for Use For the preparation of a DNA library using sub-nanogram amounts of DNA input for next generation sequencing applications
More informationMultiplexed Strand-specific RNA-Seq Library Preparation for Illumina Sequencing Platforms
Multiplexed Strand-specific RNA-Seq Library Preparation for Illumina Sequencing Platforms Important Things to know before you start: This protocol generates strand-specific reads, but may lead to slightly
More informationHigh Throughput Sequencing Technologies. UCD Genome Center Bioinformatics Core Monday 15 June 2015
High Throughput Sequencing Technologies UCD Genome Center Bioinformatics Core Monday 15 June 2015 Sequencing Explosion www.genome.gov/sequencingcosts http://t.co/ka5cvghdqo Sequencing Explosion 2011 PacBio
More informationMolecular Biology and Functional Genomic Core Facility
Molecular Biology and Functional Genomic Core Facility General Presentation Dr Odile Neyret Core Manager Myriam Rondeau Research Assistant Agnès Dumont Research Assistant Institut de recherche clinique
More informationOutline General NGS background and terms 11/14/2016 CONFLICT OF INTEREST. HLA region targeted enrichment. NGS library preparation methodologies
Eric T. Weimer, PhD, D(ABMLI) Assistant Professor, Pathology & Laboratory Medicine, UNC School of Medicine Director, Molecular Immunology Associate Director, Clinical Flow Cytometry, HLA, and Immunology
More informationHaloPlex HS. Get to Know Your DNA. Every Single Fragment. Kevin Poon, Ph.D.
HaloPlex HS Get to Know Your DNA. Every Single Fragment. Kevin Poon, Ph.D. Sr. Global Product Manager Diagnostics & Genomics Group Agilent Technologies For Research Use Only. Not for Use in Diagnostic
More informationHigh Throughput Sequencing Technologies. J Fass UCD Genome Center Bioinformatics Core Monday June 16, 2014
High Throughput Sequencing Technologies J Fass UCD Genome Center Bioinformatics Core Monday June 16, 2014 Sequencing Explosion www.genome.gov/sequencingcosts http://t.co/ka5cvghdqo Sequencing Explosion
More informationNovel methods for RNA and DNA- Seq analysis using SMART Technology. Andrew Farmer, D. Phil. Vice President, R&D Clontech Laboratories, Inc.
Novel methods for RNA and DNA- Seq analysis using SMART Technology Andrew Farmer, D. Phil. Vice President, R&D Clontech Laboratories, Inc. Agenda Enabling Single Cell RNA-Seq using SMART Technology SMART
More informationQuality Control in Illumina Sequencing Workflows Using the TapeStation System
pplication Note Genomics Quality Control in Illumina Sequencing Workflows Using the TapeStation System uthors Nicolle Diessl, Ute Ernst, ngela Schulz, and Stephan Wolf DKFZ Genomics and Proteomics Core
More informationRNA-Seq data analysis course September 7-9, 2015
RNA-Seq data analysis course September 7-9, 2015 Peter-Bram t Hoen (LUMC) Jan Oosting (LUMC) Celia van Gelder, Jacintha Valk (BioSB) Anita Remmelzwaal (LUMC) Expression profiling DNA mrna protein Comprehensive
More informationApplication Note. Genomics. Abstract. Authors. Kirill Gromadski Ruediger Salowsky Susanne Glueck Agilent Technologies Waldbronn, Germany
Improving sample quality for target enrichment and next-gen sequencing with the Agilent High Sensitivity DNA Kit and the Agilent SureSelect Target Enrichment Platform Application Note Genomics Authors
More informationSMARTer Ultra Low RNA Kit for Illumina Sequencing Two powerful technologies combine to enable sequencing with ultra-low levels of RNA
SMARTer Ultra Low RNA Kit for Illumina Sequencing Two powerful technologies combine to enable sequencing with ultra-low levels of RNA The most sensitive cdna synthesis technology, combined with next-generation
More informationA Crash Course in NGS for GI Pathologists. Sandra O Toole
A Crash Course in NGS for GI Pathologists Sandra O Toole The Sanger Technique First generation sequencing Uses dideoxynucleotides (dideoxyadenine, dideoxyguanine, etc) These are molecules that resemble
More informationComplete protocol in 110 minutes Enzymatic fragmentation without sonication One-step fragmentation/tagging to save time
Molecular Cloning Laboratories Manual Version 1.2 Product name: MCNext UT DNA Sample Prep Kit Cat #: MCUDS-4, MCUDS-24, MCUDS-96 Description: This protocol explains how to prepare up to 96 pooled indexed
More informationresequencing storage SNP ncrna metagenomics private trio de novo exome ncrna RNA DNA bioinformatics RNA-seq comparative genomics
RNA Sequencing T TM variation genetics validation SNP ncrna metagenomics private trio de novo exome mendelian ChIP-seq RNA DNA bioinformatics custom target high-throughput resequencing storage ncrna comparative
More informationBiotool DNA library prep kit V2 for Illumina
Biotool DNA library prep kit V2 for Illumina Description Biotool DNA library prep kit V2 for Illumina is developed specially for the Illumina high-throughput sequencing platform, and generates sequencing-ready
More informationThruPLEX -FD Prep Kit Instruction Manual. Single Tube Library Preparation for Illumina NGS Platforms
ThruPLEX -FD Prep Kit Instruction Manual Single Tube Library Preparation for Illumina NGS Platforms Contents Product Description... 2 Kit Contents... 2 Shipping and Storage... 2 Getting Started... 3 Input
More informationHigh Throughput Sequencing Technologies. J Fass UCD Genome Center Bioinformatics Core Monday September 15, 2014
High Throughput Sequencing Technologies J Fass UCD Genome Center Bioinformatics Core Monday September 15, 2014 Sequencing Explosion www.genome.gov/sequencingcosts http://t.co/ka5cvghdqo Sequencing Explosion
More informationRecombinant DNA Technology
History of recombinant DNA technology Recombinant DNA Technology (DNA cloning) Majid Mojarrad Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists
More informationBi 8 Lecture 4. Ellen Rothenberg 14 January Reading: from Alberts Ch. 8
Bi 8 Lecture 4 DNA approaches: How we know what we know Ellen Rothenberg 14 January 2016 Reading: from Alberts Ch. 8 Central concept: DNA or RNA polymer length as an identifying feature RNA has intrinsically
More informationIntroduction Bioo Scientific
Next Generation Sequencing Catalog 2014-2015 Introduction Bioo Scientific Bioo Scientific is a global life science company headquartered in Austin, TX, committed to providing innovative products and superior
More informationNEXTFLEX ChIP-Seq Kit (For Illumina Platforms) Catalog #NOVA (Kit contains 8 reactions) Bioo Scientific Corp V15.
NEXTFLEX ChIP-Seq Kit (For Illumina Platforms) Catalog #NOVA-5143-01 (Kit contains 8 reactions) Bioo Scientific Corp. 2015-2018 V15.07 This product is for research use only. Not for use in diagnostic procedures.
More informationGenome 373: High- Throughput DNA Sequencing. Doug Fowler
Genome 373: High- Throughput DNA Sequencing Doug Fowler Tasks give ML unity We learned about three tasks that are commonly encountered in ML Models/Algorithms Give ML Diversity Classification Regression
More informationPreparing normalized cdna libraries for transcriptome sequencing (Illumina HiSeq)
Preparing normalized cdna libraries for transcriptome sequencing (Illumina HiSeq) Last updated: Oct 28, 2016 Overview First-strand cdna is synthesized using oligo-dt containing primers and an RNA oligo
More informationNext Generation Sequencing. Tobias Österlund
Next Generation Sequencing Tobias Österlund tobiaso@chalmers.se NGS part of the course Week 4 Friday 13/2 15.15-17.00 NGS lecture 1: Introduction to NGS, alignment, assembly Week 6 Thursday 26/2 08.00-09.45
More information02 Agenda Item 03 Agenda Item
01 Agenda Item 02 Agenda Item 03 Agenda Item SOLiD 3 System: Applications Overview April 12th, 2010 Jennifer Stover Field Application Specialist - SOLiD Applications Workflow for SOLiD Application Application
More informationPerformance characteristics of the High Sensitivity DNA kit for the Agilent 2100 Bioanalyzer
Performance characteristics of the High Sensitivity DNA kit for the Agilent 2100 Bioanalyzer Technical Note 10 Measured conc. [ng/µl] 1 Y intercept = 0.09 r 2 = 0.993 0.1 0.1 1 10 Reference concentration
More informationSOLiD Total RNA-Seq Kit SOLiD RNA Barcoding Kit
SOLiD Total RNA-Seq Kit SOLiD RNA Barcoding Kit Agenda SOLiD Total RNAseq Kit Overview Kit Configurations Barcoding Kit Introduction New Small RNA and WT Workflow Small RNA Workflow Step-by-step Workflow
More informationNEXT GENERATION SEQUENCING. Innovative solutions for library
NEXT GENERATION SEQUENCING Innovative solutions for library pr NEXT GENERATION SOLUTIONS Dramatic improvements to commercial next generation sequencing (NGS) platforms have resulted in spectacular reductions
More informationObtaining DNA from degraded samples for NGS sequencing
Obtaining DNA from degraded samples for NGS sequencing A brief overview of Alexander (Sasha) Mikheyev s lecture at USC 03/13/14 Presented by Jacqueline Robinson 04/23/2014 NGS is great, but Standard protocols
More informationJetSeq DNA Library Preparation Kit. Product Manual
JetSeq DNA Library Preparation Kit Product Manual JetSeq DNA Library Preparation Kit JetSeq DNA Library Preparation Kit TABLE OF CONTENTS 1 Kit contents 04 2 Description 05 3 Storage 06 4 Safety information
More informationCM581A2: NEXT GENERATION SEQUENCING PLATFORMS AND LIBRARY GENERATION
CM581A2: NEXT GENERATION SEQUENCING PLATFORMS AND LIBRARY GENERATION Fall 2015 Instructors: Coordinator: Carol Wilusz, Associate Professor MIP, CMB Instructor: Dan Sloan, Assistant Professor, Biology,
More informationGenetics and Genomics in Medicine Chapter 3. Questions & Answers
Genetics and Genomics in Medicine Chapter 3 Multiple Choice Questions Questions & Answers Question 3.1 Which of the following statements, if any, is false? a) Amplifying DNA means making many identical
More informationBIOO LIFE SCIENCE PRODUCTS
BIOO LIFE SCIENCE PRODUCTS FOR REFERENCE PURPOSES This manual is for Reference Purposes Only. DO NOT use this protocol to run your assays. Periodically, optimizations and revisions are made to the kit
More informationTruSeq ChIP Sample Preparation
FOR RESEARCH USE ONLY Date: Illumina Kit Description: NOTE Unless familiar with the protocol in the latest version of the TruSeq ChIP Sample Preparation Guide (part # 15023092), new or less experienced
More informationGENOTYPING-BY-SEQUENCING USING CUSTOM ION AMPLISEQ TECHNOLOGY AS A TOOL FOR GENOMIC SELECTION IN ATLANTIC SALMON
GENOTYPING-BY-SEQUENCING USING CUSTOM ION AMPLISEQ TECHNOLOGY AS A TOOL FOR GENOMIC SELECTION IN ATLANTIC SALMON Matthew Baranski, Casey Jowdy, Hooman Moghadam, Ashie Norris, Håvard Bakke, Anna Sonesson,
More informationWhole genome Bisulfite Sequencing for Methylation Analysis Preparing Samples for the Illumina Sequencing Platform
Whole genome Bisulfite Sequencing for Methylation Analysis Preparing Samples for the Illumina Sequencing Platform Introduction, 2 Sample Prep Workflow, 3 Best Practices, 4 DNA Input Recommendations, 6
More informationKAPA Library Preparation Kits
Technical Data Sheet KAPA Library Preparation Kits Illumina series Product Description The KAPA Library Preparation Kit provides all of the enzymes and reaction buffers required for constructing libraries
More informationACCEL-NGS 2S DNA LIBRARY KITS
ACCEL-NGS 2S DNA LIBRARY KITS Accel-NGS 2S DNA Library Kits produce high quality libraries with an all-inclusive, easy-to-use format. The kits contain all reagents necessary to build high complexity libraries
More informationMate-pair library data improves genome assembly
De Novo Sequencing on the Ion Torrent PGM APPLICATION NOTE Mate-pair library data improves genome assembly Highly accurate PGM data allows for de Novo Sequencing and Assembly For a draft assembly, generate
More informationRequirements for Illumina Sequencing: Constructed Library Submission
Requirements for Illumina Sequencing: Constructed Library Submission Genome Sciences Centre, BC Cancer January 2018 Table of Contents Library Construction Method for DNA...1 Library Construction Method
More informationSequencing technologies. Jose Blanca COMAV institute bioinf.comav.upv.es
Sequencing technologies Jose Blanca COMAV institute bioinf.comav.upv.es Outline Sequencing technologies: Sanger 2nd generation sequencing: 3er generation sequencing: 454 Illumina SOLiD Ion Torrent PacBio
More informationMHC Region. MHC expression: Class I: All nucleated cells and platelets Class II: Antigen presenting cells
DNA based HLA typing methods By: Yadollah Shakiba, MD, PhD MHC Region MHC expression: Class I: All nucleated cells and platelets Class II: Antigen presenting cells Nomenclature of HLA Alleles Assigned
More informationTemplate Preparation FIND MEANING IN COMPLEXITY. Copyright 2014 by Pacific Biosciences of California, Inc. All rights reserved.
Template Preparation FIND MEANING IN COMPLEXITY Copyright 2014 by Pacific Biosciences of California, Inc. All rights reserved. PN 100-336-800-02 Specifics of SMRT Sequencing Data Steps of Overview SMRTbell
More informationNEBNext. for Ion Torrent LIBRARY PREPARATION KITS
NEBNext for Ion Torrent LIBRARY PREPARATION KITS NEBNEXT PRODUCTS FOR ION TORRENT Table of Contents 3 General Introduction TOOLS & RESOURCES Visit NEBNext.com to find: T he full list of products available
More informationDesign. Construction. Characterization
Design Construction Characterization DNA mrna (messenger) A C C transcription translation C A C protein His A T G C T A C G Plasmids replicon copy number incompatibility selection marker origin of replication
More informationSample Quality Control in Agilent NGS Solutions
pplication Note Next Generation Sequencing Sample Quality Control in gilent NGS Solutions uthors Madhurima iswas and Shweta Sharma gilent Technologies, Inc. La Jolla, C US Weiwei Liu, Tracy Liu, and David
More informationNext Generation Sequencing. Target Enrichment
Next Generation Sequencing Target Enrichment Next Generation Sequencing Your Partner in Every Step from Sample to Data NGS: Revolutionizing Genetic Analysis with Single-Molecule Resolution Next generation
More informationNGS Library Construction Kit User Guide
NGS Library Construction Kit User Guide Catalog Number BX2000-08M REV. 1.0 11.21.16 Part number 40023 LEGAL NOTICES Technical Services Limited Use Label License Limited Warranty Trademark Information Regulatory
More informationTargeted Sequencing Using Droplet-Based Microfluidics. Keith Brown Director, Sales
Targeted Sequencing Using Droplet-Based Microfluidics Keith Brown Director, Sales brownk@raindancetech.com Who we are: is a Provider of Microdroplet-based Solutions The Company s RainStorm TM Technology
More information