Appendix I: Brief Unix Reference from Now On, We Assume That You Have Access to a Unix Terminal Window

Total Page:16

File Type:pdf, Size:1020Kb

Appendix I: Brief Unix Reference from Now On, We Assume That You Have Access to a Unix Terminal Window Appendix I: Brief Unix reference From now on, we assume that you have access to a Unix terminal window. We will be focusing on operations that are possible at the command line, as this is where you are most effective in the long run. Operating systems File system Whenever you are using a computer you interact with it with the help of an operating system (OS), a vital interface between hardware and user. The operating system does a Examples of directories and their contents number of different things. For instance, multiple programs are often run at the same time and in this situation the operating system allocates resources to the different programs or / root of file system may be able to appropriately interrupt programs. Another common feature of an /bin executable binary files operating system is a graphical user interface, originally developed for personal /dev special files used to represent real physical devices computers. Examples of popular operating systems are Microsoft Windows, Mac OSX /etc commands and files used for system administration and Linux. /home contains a home directory for each user of the system /home/joe home directory of user joe Linux is an example of a Unix (or "Unix-like") operating system. Unix was originally /lib libraries used by various programs and programming languages developed in 1969 at Bell Laboratories in the US. Many different flavours of the Unix a "scratch" area where any user can store files temporarily operating systems have been developed, such as Solaris, HP-UX and AIX and there is a /tmp system files and directories that you share with other users number of freely available Unix or Unix-like systems such as GNU/Linux in different /usr distributions such as Red Hat Enterprise Linux, Fedora, SUSE Linux Enterprise, openSUSE and Ubuntu. Moving around in the file system If you are at a personal computer your access to a Unix depends on what operating system you are using. For instance, Microsoft Windows is not based on Unix and does Find out what directory you are currently in: not provide a Unix interface. If you would like to have a Unix environment within Windows a possible choice is to install Cygwin (http://www.cygwin.com). Cygwin has a % pwd "present working directory" lot of Unix functionality and useful Unix programs and it also includes Perl (see also the Appendix III). The Mac OS X operating system is based on Unix and all you need to do Move to a specific directory with cd, "change directory": to communicate with Unix commands is to open a terminal window (available under Applications-Utilities). Same thing if you are at a computer with Linux or any other Unix % cd /tmp change directory to /tmp flavour; just open a terminal window to get started. % cd .. go up one level in directory tree % cd (without argument) go to your home directory Accessing a UNIX computer Find out what files are in the current directory: Even though you do not have Unix at your personal computer, you can connect to a Unix-based server and operate from there. In fact, this is a common mode of working in % ls show files in current directory bioinformatics. You typically make use of an SSH (Secure Shell) client program to % ls -al show files in current directory connect. SSH is a network protocol to allow data to be transferred between two networked computers. The SSH client program communicates with a SSH daemon The ls -al command will result in a more detailed output, as in this example: running on the server side. A typical application of the SSH client program is to login to a remote computer and execute commands at the remote computer. Examples of freely -rw-r--r-- 1 joe users 383269 2007-11-25 16:54 PF02854.txt drwxr-xr-x 9 joe users 656 2003-04-04 20:12 scripts/ available implementations of SSH are openSSH (http://www.openssh.com/), copSSH -rw-r--r-- 1 joe users 4898 2006-09-12 09:12 README.txt (http://www.itefix.no/i2/copssh) and PuTTY -rwxr-xr-x 1 joe users 120635 2004-08-03 01:47 dnapars* (http://www.chiark.greenend.org.uk/~sgtatham/putty/). In this listing there are in each line a set of characters describing the file status. In a string Removing files like -rw-r--r--, the first '- ' means that it is a regular file ( a 'd' says that it is a directory). The next symbols are three groups of three where the first is what the owner % rm seq.fa can do, second what the group members can do and third what the other users can do. In each group of three, the symbols are 'r' = readable, 'w' = writable, 'x' = executable. To Creating and removing directories change the file attributes, see documentation on the Unix command chmod. % mkdir dirname As with many other Unix commands, the wildcard * may be used with ls. The following % rmdir dirname command will list files with names beginning with 'HIV': Viewing and editing files % ls HIV* % cat seq.fa (will show the contents of seq.fa on the screen) Manipulating Files and Directories The cat command may also be used to merge (concatenate) files. This command will merge three different files, file1, file2 and file3 into a new file newfile: Copying files % cat file1 file2 file3 > newfile cp [source] [destination] The symbol > means that we are redirecting the output of the cat program to a file Examples: instead of the standard output (which is the screen). We can also append to an existing file, using the symbols >>: 1) Copy the file /tmp/seq.fa to the current working directory. The current directory is represented by a dot '.': % cat file4 >> newfile % cp /tmp/seq.fa . Viewing a text file on the screen one page at a time. 2) Copy the file /tmp/seq.fa to another file in /tmp named seq2.fa: To view a text file on the screen, use either more or less. With less: % cp /tmp/seq.fa /tmp/seq2.fa % less seq.fa 3) Copy all files (*) in the directory /home/joe/seqfiles to the directory /tmp: Some useful keys for less are: % cp /home/joe/seqfiles/* /tmp space : move down one page Moving and renaming files enter : go down one line u : go up (back) mv [source] [destination] /HIV : search for 'HIV' q : quit program Examples: Viewing or extracting the first or last lines of a file 1) Rename the file seq.fa in /tmp to seq2.fa: % mv /tmp/seq.fa /tmp/seq2.fa First and last lines of a file may be displayed using the commands head and tail, respectively. 2) Move all files (*) in the directory /home/joe/seqfiles to the directory /tmp: % head seq.fa (by default head will show the first 10 lines of the file) % mv /home/joe/seqfiles/* /tmp % head -1000 seq.fa (first 1000 lines of file will be extracted) % cut -f1,2 -d ';' dat2.txt Text-mode editor vi The output will be: % vi seq.fa A;2 B;5 The editor vi is very useful whenever you do not have access to a graphical editor. F;4 Description of it, however, requires a book on its own. For more information on vi the reader is referred to other sources such as: Sorting http://en.wikibooks.org/wiki/Learning_the_vi_Editor http://unixhelp.ed.ac.uk/vi/ Consider the file dat3.txt that contains: Graphical editors A 12 1300 1306 C 11 1500 1458 B 17 1620 1700 Examples of graphical editors are emacs (http://www.gnu.org/software/emacs/), gedit (http://www.gedit.org) and nedit (http://www.nedit.org). The lines may be sorted using sort: % sort dat3.txt Extracting file components with cut The output will be: Consider the content of a file dat.txt where the columns are separated with tabs: A 12 1300 1306 1 12 1300 1306 B 17 1620 1700 2 11 1500 1458 C 11 1500 1458 3 17 1620 1700 The sort utility sorts lines alphabetically by default. Sorting is done numerically if we use the option -n. In addition, we may specify sorting with respect to a specific column, We may extract the columns 1 and 3 with cut: using the parameter -k: % cut -f1,3 dat.txt % sort -n -k2 dat3.txt which produces: The output is then: 1 1300 C 11 1500 1458 2 1500 A 12 1300 1306 3 1620 B 17 1620 1700 The fields or columns to be extracted are specified with the -f option. The default Note that now the values in column 2 are in numerical order. We may also reverse the separator is tab, but we may use any separator. The separator is specified with the -d order of sorting with the -r parameter: option to cut. Consider the file dat2.txt which contains: % sort -n -k2 -r dat3.txt A;2;4500 B;5;4505 F;4;4510 Unique lines We try the cut command: The uniq command is used to identify the unique lines in a file: A highly useful Unix utility is the grep command, used to search files for text strings or % uniq sortedfile regular expression matching: For this to work well the lines in the file need to be sorted first with sort. A useful option % grep ">" seq.fa | wc to uniq is -c . The effect is to list the number of times each line occurs: In this example grep will identify all lines in seq.fa that contain '>'. The output of grep % uniq -c sortedfile will then be directed to wc.
Recommended publications
  • Administering Unidata on UNIX Platforms
    C:\Program Files\Adobe\FrameMaker8\UniData 7.2\7.2rebranded\ADMINUNIX\ADMINUNIXTITLE.fm March 5, 2010 1:34 pm Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta UniData Administering UniData on UNIX Platforms UDT-720-ADMU-1 C:\Program Files\Adobe\FrameMaker8\UniData 7.2\7.2rebranded\ADMINUNIX\ADMINUNIXTITLE.fm March 5, 2010 1:34 pm Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Beta Notices Edition Publication date: July, 2008 Book number: UDT-720-ADMU-1 Product version: UniData 7.2 Copyright © Rocket Software, Inc. 1988-2010. All Rights Reserved. Trademarks The following trademarks appear in this publication: Trademark Trademark Owner Rocket Software™ Rocket Software, Inc. Dynamic Connect® Rocket Software, Inc. RedBack® Rocket Software, Inc. SystemBuilder™ Rocket Software, Inc. UniData® Rocket Software, Inc. UniVerse™ Rocket Software, Inc. U2™ Rocket Software, Inc. U2.NET™ Rocket Software, Inc. U2 Web Development Environment™ Rocket Software, Inc. wIntegrate® Rocket Software, Inc. Microsoft® .NET Microsoft Corporation Microsoft® Office Excel®, Outlook®, Word Microsoft Corporation Windows® Microsoft Corporation Windows® 7 Microsoft Corporation Windows Vista® Microsoft Corporation Java™ and all Java-based trademarks and logos Sun Microsystems, Inc. UNIX® X/Open Company Limited ii SB/XA Getting Started The above trademarks are property of the specified companies in the United States, other countries, or both. All other products or services mentioned in this document may be covered by the trademarks, service marks, or product names as designated by the companies who own or market them. License agreement This software and the associated documentation are proprietary and confidential to Rocket Software, Inc., are furnished under license, and may be used and copied only in accordance with the terms of such license and with the inclusion of the copyright notice.
    [Show full text]
  • Types and Programming Languages by Benjamin C
    < Free Open Study > . .Types and Programming Languages by Benjamin C. Pierce ISBN:0262162091 The MIT Press © 2002 (623 pages) This thorough type-systems reference examines theory, pragmatics, implementation, and more Table of Contents Types and Programming Languages Preface Chapter 1 - Introduction Chapter 2 - Mathematical Preliminaries Part I - Untyped Systems Chapter 3 - Untyped Arithmetic Expressions Chapter 4 - An ML Implementation of Arithmetic Expressions Chapter 5 - The Untyped Lambda-Calculus Chapter 6 - Nameless Representation of Terms Chapter 7 - An ML Implementation of the Lambda-Calculus Part II - Simple Types Chapter 8 - Typed Arithmetic Expressions Chapter 9 - Simply Typed Lambda-Calculus Chapter 10 - An ML Implementation of Simple Types Chapter 11 - Simple Extensions Chapter 12 - Normalization Chapter 13 - References Chapter 14 - Exceptions Part III - Subtyping Chapter 15 - Subtyping Chapter 16 - Metatheory of Subtyping Chapter 17 - An ML Implementation of Subtyping Chapter 18 - Case Study: Imperative Objects Chapter 19 - Case Study: Featherweight Java Part IV - Recursive Types Chapter 20 - Recursive Types Chapter 21 - Metatheory of Recursive Types Part V - Polymorphism Chapter 22 - Type Reconstruction Chapter 23 - Universal Types Chapter 24 - Existential Types Chapter 25 - An ML Implementation of System F Chapter 26 - Bounded Quantification Chapter 27 - Case Study: Imperative Objects, Redux Chapter 28 - Metatheory of Bounded Quantification Part VI - Higher-Order Systems Chapter 29 - Type Operators and Kinding Chapter 30 - Higher-Order Polymorphism Chapter 31 - Higher-Order Subtyping Chapter 32 - Case Study: Purely Functional Objects Part VII - Appendices Appendix A - Solutions to Selected Exercises Appendix B - Notational Conventions References Index List of Figures < Free Open Study > < Free Open Study > Back Cover A type system is a syntactic method for automatically checking the absence of certain erroneous behaviors by classifying program phrases according to the kinds of values they compute.
    [Show full text]
  • UNIX Workshop Series: Quick-Start Objectives
    Part I UNIX Workshop Series: Quick-Start Objectives Overview – Connecting with ssh Command Window Anatomy Command Structure Command Examples Getting Help Files and Directories Wildcards, Redirection and Pipe Create and edit files Overview Connecting with ssh Open a Terminal program Mac: Applications > Utilities > Terminal ssh –Y [email protected] Linux: In local shell ssh –Y [email protected] Windows: Start Xming and PuTTY Create a saved session for the remote host name centos.css.udel.edu using username Connecting with ssh First time you connect Unix Basics Multi-user Case-sensitive Bash shell, command-line Commands Command Window Anatomy Title bar Click in the title bar to bring the window to the front and make it active. Command Window Anatomy Login banner Appears as the first line of a login shell. Command Window Anatomy Prompts Appears at the beginning of a line and usually ends in $. Command Window Anatomy Command input Place to type commands, which may have options and/or arguments. Command Window Anatomy Command output Place for command response, which may be many lines long. Command Window Anatomy Input cursor Typed text will appear at the cursor location. Command Window Anatomy Scroll Bar Will appear as needed when there are more lines than fit in the window. Command Window Anatomy Resize Handle Use the mouse to change the window size from the default 80x24. Command Structure command [arguments] Commands are made up of the actual command and its arguments. command -options [arguments] The arguments are further broken down into the command options which are single letters prefixed by a “-” and other arguments that identify data for the command.
    [Show full text]
  • State Notation Language and Sequencer Users' Guide
    State Notation Language and Sequencer Users’ Guide Release 2.0.99 William Lupton ([email protected]) Benjamin Franksen ([email protected]) June 16, 2010 CONTENTS 1 Introduction 3 1.1 About.................................................3 1.2 Acknowledgements.........................................3 1.3 Copyright...............................................3 1.4 Note on Versions...........................................4 1.5 Notes on Release 2.1.........................................4 1.6 Notes on Releases 2.0.0 to 2.0.12..................................7 1.7 Notes on Release 2.0.........................................9 1.8 Notes on Release 1.9......................................... 12 2 Installation 13 2.1 Prerequisites............................................. 13 2.2 Download............................................... 13 2.3 Unpack................................................ 14 2.4 Configure and Build......................................... 14 2.5 Building the Manual......................................... 14 2.6 Test.................................................. 15 2.7 Use.................................................. 16 2.8 Report Bugs............................................. 16 2.9 Contribute.............................................. 16 3 Tutorial 19 3.1 The State Transition Diagram.................................... 19 3.2 Elements of the State Notation Language.............................. 19 3.3 A Complete State Program...................................... 20 3.4 Adding a
    [Show full text]
  • System Log Commands
    System Log Commands • system set-log , on page 2 • show system logging-level, on page 3 • show log, on page 4 System Log Commands 1 System Log Commands system set-log system set-log To set the log level and log type of messages, use the system set-log command in privileged EXEC mode. system set-log level {debug | info | warning | error | critical} logtype {configuration | operational | all} Syntax Description level Specifies the log level. debug Logs all messages. info Logs all messages that have info and higher severity level. warning Logs all messages that have warning and higher severity level. error Logs all messages that have error and higher severity level. critical Logs all messages that have critical severity level. logtype Specifies the log type. configuration Configuration log messages are recorded. operational Operational log messages are recorded. all All types of log messages are recorded. Command Default For the configuration log, info is the default level. For the operational log, warning is the default level. Command Modes Privileged EXEC (#) Command History Release Modification 3.5.1 This command was introduced. Usage Guidelines After a system reboot, the modified logging configuration is reset to the default level, that is, info for the configuration log and warning for the operational log. Example The following example shows how to configure a log level: nfvis# system set-log level error logtype all System Log Commands 2 System Log Commands show system logging-level show system logging-level To view the log level and log type settings, use the show system logging-level command in privileged EXEC mode.
    [Show full text]
  • A Rapid, Sensitive, Scalable Method for Precision Run-On Sequencing
    bioRxiv preprint doi: https://doi.org/10.1101/2020.05.18.102277; this version posted May 19, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 A rapid, sensitive, scalable method for 2 Precision Run-On sequencing (PRO-seq) 3 4 Julius Judd1*, Luke A. Wojenski2*, Lauren M. Wainman2*, Nathaniel D. Tippens1, Edward J. 5 Rice3, Alexis Dziubek1,2, Geno J. Villafano2, Erin M. Wissink1, Philip Versluis1, Lina Bagepalli1, 6 Sagar R. Shah1, Dig B. Mahat1#a, Jacob M. Tome1#b, Charles G. Danko3,4, John T. Lis1†, 7 Leighton J. Core2† 8 9 *Equal contribution 10 1Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA 11 2Department of Molecular and Cell Biology, Institute of Systems Genomics, University of 12 Connecticut, Storrs, CT 06269, USA 13 3Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, 14 NY 14853, USA 15 4Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, 16 Ithaca, NY 14853, USA 17 #aPresent address: Koch Institute for Integrative Cancer Research, Massachusetts Institute of 18 Technology, Cambridge, MA 02139, USA 19 #bPresent address: Department of Genome Sciences, University of Washington, Seattle, WA, 20 USA 21 †Correspondence to: [email protected] or [email protected] 22 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.05.18.102277; this version posted May 19, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Dell EMC Powerstore CLI Guide
    Dell EMC PowerStore CLI Guide May 2020 Rev. A01 Notes, cautions, and warnings NOTE: A NOTE indicates important information that helps you make better use of your product. CAUTION: A CAUTION indicates either potential damage to hardware or loss of data and tells you how to avoid the problem. WARNING: A WARNING indicates a potential for property damage, personal injury, or death. © 2020 Dell Inc. or its subsidiaries. All rights reserved. Dell, EMC, and other trademarks are trademarks of Dell Inc. or its subsidiaries. Other trademarks may be trademarks of their respective owners. Contents Additional Resources.......................................................................................................................4 Chapter 1: Introduction................................................................................................................... 5 Overview.................................................................................................................................................................................5 Use PowerStore CLI in scripts.......................................................................................................................................5 Set up the PowerStore CLI client........................................................................................................................................5 Install the PowerStore CLI client..................................................................................................................................
    [Show full text]
  • Specification of the "Sequ" Command
    1 Specification of the "sequ" command 2 Copyright © 2013 Bart Massey 3 Revision 0, 1 October 2013 4 This specification describes the "universal sequence" command sequ. The sequ command is a 5 backward-compatible set of extensions to the UNIX [seq] 6 (http://www.gnu.org/software/coreutils/manual/html_node/seq-invo 7 cation.html) command. There are many implementations of seq out there: this 8 specification is built on the seq supplied with GNU Coreutils version 8.21. 9 The seq command emits a monotonically increasing sequence of numbers. It is most commonly 10 used in shell scripting: 11 TOTAL=0 12 for i in `seq 1 10` 13 do 14 TOTAL=`expr $i + $TOTAL` 15 done 16 echo $TOTAL 17 prints 55 on standard output. The full sequ command does this basic counting operation, plus 18 much more. 19 This specification of sequ is in several stages, known as compliance levels. Each compliance 20 level adds required functionality to the sequ specification. Level 1 compliance is equivalent to 21 the Coreutils seq command. 22 The usual specification language applies to this document: MAY, SHOULD, MUST (and their 23 negations) are used in the standard fashion. 24 Wherever the specification indicates an error, a conforming sequ implementation MUST 25 immediately issue appropriate error message specific to the problem. The implementation then 26 MUST exit, with a status indicating failure to the invoking process or system. On UNIX systems, 27 the error MUST be indicated by exiting with status code 1. 28 When a conforming sequ implementation successfully completes its output, it MUST 29 immediately exit, with a status indicating success to the invoking process or systems.
    [Show full text]
  • Epmp Command Line Interface User Guide
    USER GUIDE ePMP Command Line Interface ePMP Command Line Interface User Manual Table of Contents 1 Introduction ...................................................................................................................................... 3 1.1 Purpose ................................................................................................................................ 3 1.2 Command Line Access ........................................................................................................ 3 1.3 Command usage syntax ...................................................................................................... 3 1.4 Basic information ................................................................................................................. 3 1.4.1 Context sensitive help .......................................................................................................... 3 1.4.2 Auto-completion ................................................................................................................... 3 1.4.3 Movement keys .................................................................................................................... 3 1.4.4 Deletion keys ....................................................................................................................... 4 1.4.5 Escape sequences .............................................................................................................. 4 2 Command Line Interface Overview ..............................................................................................
    [Show full text]
  • Powerview Command Reference
    PowerView Command Reference TRACE32 Online Help TRACE32 Directory TRACE32 Index TRACE32 Documents ...................................................................................................................... PowerView User Interface ............................................................................................................ PowerView Command Reference .............................................................................................1 History ...................................................................................................................................... 12 ABORT ...................................................................................................................................... 13 ABORT Abort driver program 13 AREA ........................................................................................................................................ 14 AREA Message windows 14 AREA.CLEAR Clear area 15 AREA.CLOSE Close output file 15 AREA.Create Create or modify message area 16 AREA.Delete Delete message area 17 AREA.List Display a detailed list off all message areas 18 AREA.OPEN Open output file 20 AREA.PIPE Redirect area to stdout 21 AREA.RESet Reset areas 21 AREA.SAVE Save AREA window contents to file 21 AREA.Select Select area 22 AREA.STDERR Redirect area to stderr 23 AREA.STDOUT Redirect area to stdout 23 AREA.view Display message area in AREA window 24 AutoSTOre ..............................................................................................................................
    [Show full text]
  • Unix: Beyond the Basics
    Unix: Beyond the Basics BaRC Hot Topics – September, 2018 Bioinformatics and Research Computing Whitehead Institute http://barc.wi.mit.edu/hot_topics/ Logging in to our Unix server • Our main server is called tak4 • Request a tak4 account: http://iona.wi.mit.edu/bio/software/unix/bioinfoaccount.php • Logging in from Windows Ø PuTTY for ssh Ø Xming for graphical display [optional] • Logging in from Mac ØAccess the Terminal: Go è Utilities è Terminal ØXQuartz needed for X-windows for newer OS X. 2 Log in using secure shell ssh –Y user@tak4 PuTTY on Windows Terminal on Macs Command prompt user@tak4 ~$ 3 Hot Topics website: http://barc.wi.mit.edu/education/hot_topics/ • Create a directory for the exercises and use it as your working directory $ cd /nfs/BaRC_training $ mkdir john_doe $ cd john_doe • Copy all files into your working directory $ cp -r /nfs/BaRC_training/UnixII/* . • You should have the files below in your working directory: – foo.txt, sample1.txt, exercise.txt, datasets folder – You can check they’re there with the ‘ls’ command 4 Unix Review: Commands Ø command [arg1 arg2 … ] [input1 input2 … ] $ sort -k2,3nr foo.tab -n or -g: -n is recommended, except for scientific notation or start end a leading '+' -r: reverse order $ cut -f1,5 foo.tab $ cut -f1-5 foo.tab -f: select only these fields -f1,5: select 1st and 5th fields -f1-5: select 1st, 2nd, 3rd, 4th, and 5th fields $ wc -l foo.txt How many lines are in this file? 5 Unix Review: Common Mistakes • Case sensitive cd /nfs/Barc_Public vs cd /nfs/BaRC_Public -bash: cd: /nfs/Barc_Public:
    [Show full text]
  • Transcriptome-Wide M a Detection with DART-Seq
    Transcriptome-wide m6A detection with DART-Seq Kate D. Meyer ( [email protected] ) Duke University School of Medicine https://orcid.org/0000-0001-7197-4054 Method Article Keywords: RNA, epitranscriptome, RNA methylation, m6A, methyladenosine, DART-Seq Posted Date: September 23rd, 2019 DOI: https://doi.org/10.21203/rs.2.12189/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/7 Abstract m6A is the most abundant internal mRNA modication and plays diverse roles in gene expression regulation. Much of our current knowledge about m6A has been driven by recent advances in the ability to detect this mark transcriptome-wide. Antibody-based approaches have been the method of choice for global m6A mapping studies. These methods rely on m6A antibodies to immunoprecipitate methylated RNAs, followed by next-generation sequencing to identify m6A-containing transcripts1,2. While these methods enabled the rst identication of m6A sites transcriptome-wide and have dramatically improved our ability to study m6A, they suffer from several limitations. These include requirements for high amounts of input RNA, costly and time-consuming library preparation, high variability across studies, and m6A antibody cross-reactivity with other modications. Here, we describe DART-Seq (deamination adjacent to RNA modication targets), an antibody-free method for global m6A detection. In DART-Seq, the C to U deaminating enzyme, APOBEC1, is fused to the m6A-binding YTH domain. This fusion protein is then introduced to cellular RNA either through overexpression in cells or with in vitro assays, and subsequent deamination of m6A-adjacent cytidines is then detected by RNA sequencing to identify m6A sites.
    [Show full text]