7/23/2023 0 Comments Solarwinds server u![]() Start the Serv-U instance and import the Server Identity, created in Step 3 above.If required, enter the listener IP addresses and other host-specific Serv-U settings for this specific machine. Stop the installed Serv-U instance and copy the definition, created in Step 2 above, into the Serv-U program data directory.And will be used to replicate all other Serv-U instances that compose the entire Server.įor each subsequent machine on which you want to install an instance of the server, Install Serv-U on the machine. This comprises a number of files located in the Serv-U program data directory including Serv-U.Archive. and create a movable copy of the server definition. The complexity requirements for the password are displayed when you click in the password field.Ĭlose this instance of Serv-U.Enter a password and confirm, then click Export.This creates the server definition that will be used by all the subsequent instances of this server. Install Serv-U MFT 15.3.2 on a clean host machine.Ī Server Identity is automatically created.Ĭonfigure the server as required. Only the Serv-U Local Administrator or full privileged System Administrators can export and import a server Identity. If you need to set up multiple instances of your server across host machines - for example, for load balancing, this can be done as follows: Set up a new multi-tier Serv-U Server across multiple machines If you are replicating an existing server identity for use with multiple instances in a multi-tier server set-up, see the following section.Ĭlick Import to launch the Import Server Identity pop-up.Ĭlick Browse and navigate to the location containing the Server Identity. ![]() If you need to import an existing server identity - for example, if you have moved the location of the server: Only the Serv-U Local Administrator or full privileged System Administrators can import a Server Identity. Store the export file and password in a safe, secured location. The complexity requirements for the password are displayed when you click in the password field. Navigate to Global > Server Details > Server Identity.Ĭlick Export to launch the Export Server Identity pop-up.Ĭlick Browse and navigate to the directory into which you want to copy the Server Identity.Įnter the file name for the Server Identity and click Select.Įnter a password and confirm, and then click Export. Only the Serv-U Local Administrator or full privileged System Administrators can export a server Identity. Once you have set up the Serv-U 15.3.2 or later File Server, you can export and save the Server Identity to use for further instances of this server - for example, if you are creating a multi-tier file server for load balancing or if you need to move the server to another machine. ![]() Navigate to Global > Server Details > Server Identity.If you are required to create a new Server Identifier (rather than import an existing identifier): You can confirm this if in doubt by navigating to Global > Server Details > Server Identity. If you install a copy of Serv-U 15.3.2 or later for the first time and do not see the following message then a server identifier has been automatically created. Creating, exporting, and importing the Server Identity Create a server identity
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7/23/2023 0 Comments Pro tools requirements![]() or plugging your computer into a speaker system. the computer’s built in speakers (which provide no bass, so this is essentially useless) 2. So you have three playback options here: 1. You could also plug in speakers, using a 3.5mm -> RCA cable for speakers (or an amp) with RCA inputs or a 3.5mm -> 1/4″ Jack Plug cable to plug into any studio monitors that have a 1.4″ TRS Jack input, which is most of them. ![]() The Mac’s internal soundcard – called “BUILT-IN OUTPUT” in Pro Tools Playback Engine – will pump the sound either out through the Mac’s stereo speakers, or to the headphone port when you plug in headphones. The image above shows a typical MacBook Pro with a headphone port. Record MIDI using a USB MIDI controller (basically a keyboard).Play back MIDI and hear it from your computer’s internal soundcard.Arrange samples or Loops on your timeline, and hear them from your computer’s internal soundcard.Play back anything on your timeline using your computers internal soundcard.Not having an audio interface doesn’t make using Pro Tools impossible. What can I do in Pro Tools without an Audio Interface? To make a decision on which audio interface to buy, we have a bsic buying guide right here. So if you need or want to record something using a jack plug or a microphone, you’ll need something like this. This means real vocals, guitars, drums, saxophones, etc. Without an Audio Interface like this, you wouldn’t be able to record anything that requires a microphone or a jack cable. You’ll also see a headphone output on the back as well. Have a look at the Presonus 1810 above ( if you’ve read our other articles, you’ll know we like this one!), you’ll see Microphone inputs on the front and Line outs on the back – these are where you’d plug your speakers into. Converts your analog audio to digital data.Provides Microphone, Jack and Instrument Inputs.To over simplify – an audio interface does 3 main things: So let’s look at Audio Interfaces for a second. Because you can use your DAW without an audio interface and you can record without an audio interface, but you’re seriously limited in what you can do – might that actually might be ok!! The usual answers “yes”, and “if you want to record, you will” appear, but even those can be confusing answers because there’s no context. The question of ‘do I need an audio interface’ pops up every so often on the forums. So if you’re still trying to figure it out – don’t worry! It’s complicated and we’re here to try and make it a little easier. But at it’s core – any kind of music production is ‘audio engineering’ and that, by definition, is difficult. The ability to make production music at home has been made so much easier in the last 20 years with the very broad commercialisation of equipment and wide availability of software. Roland Rubix 44 Interface Do I need an interface? 7/23/2023 0 Comments Go shuttle newark airportLet us do the driving so you can spend more time having fun instead of driving around looking for a parking spot. Just tell us where you would like to be dropped off and where you would like to be picked up. Broadway Shows, Saks Fifth Avenue Shopping, Museum, New World Trade Center and so much more. SUSSEXPRESS provides transportation to NY. SUSSEXPRESS provides service for private events such as corporate events, weddings, golf outings, wine tastings, family outings, and private parties. Services to Manhattan NYC Cruise Ports, Brooklyn NY Cruise Ports, and Cape Liberty Cruise Port Bayonne NJ. SUSSEXPRESS provides service to all cruise ports. Services to Newark, JFK, LaGuardia, Stewart International, Westchester County Airport, Trenton NJ airports. SUSSEXPRESS provides service to all airports. Based in Hamburg, New Jersey, we proudly serve customers traveling throughout the Tri-State area. Reach out to us today to discuss our highly competitive rates and to book your rides.Ĭontact us to request our affordable service. And if it’s your wedding day, you can rest assured that you and your party will experience pleasant, on-schedule transportation to and from your hotel and venue. We’ll get your group to conferences, dinner meetings, golf outings, wine tastings, and more on time. ![]() Talk to us when you need to arrange travel for your clients or employees. If you are trying to get to the local cruise ports or to Dover Train Station, Newark Penn Station, or Wayne Transit Center, we’ve got you covered.Īdditionally, we offer private shuttling services for corporate and private events. Additionally, we offer rides to all of New York City’s attractions, as well as to Jets, Giants, and Devils games, and to concerts at local venues. We provide service to major shopping malls and retail outlets. Shuttle Services for Entertainment and Private EventsĮnjoy your day of leisure by riding in style to your destination. Currently, we do not upcharge for early morning or late night rides. Once we receive your flight number, we monitor incoming flights for delays and early arrivals. Reliable Airport Shuttle Serviceįrom dropping you off at the gate on time to picking you up after you come home from your trip, our services are the dependable and pleasant way to make an airport commute. ![]() We ask that all trips be booked 48 hours in advance, and our online booking system is accessible 24/7. Call us today to schedule your next ride. Whether you’re trying to catch a flight at LaGuardia Airport or arrive for a reservation in the heart of New York City, our company of safe and punctual drivers gets you where you need to be. Our drivers carry passengers to the Tri-State area’s major airports and attractions, and we also cater to private events. Convenient Transportation Services in Sussex County, NJįor comfortable and convenient travel to and from your destination, call SUSSEXPRESS for transportation services in Hamburg, NJ. 7/23/2023 0 Comments Alpha protocol torrent![]() PS: The core aspect of this game is the atmosphere. The game is divided into 15 parts which take about 5-10 minutes each the progress is saved at their beginnings. The music is revisited (with a single exception) with a selection of more atmospheric tracks. A highly talented voice actor has recorded the full narration of the game. The environments are made from scratch, the story is altered and transformed into poem form. GENIE is the re-creation of Drizzlepath: Genie which was released in 2016. After that is done, the next poem-spot reveals, and so on until the next level begins. When that happens, the camera view changes and the narration begins. When the player stands on them, they have to find a light source around them and focus on it. When they collect all the shards, the poem-spots start to appear one by one. In the first half of the levels, the player collects crystal shards scattered around the map. A rich story that’s presented in a poetic form combined with immersive graphics, fantasy style, gorgeous natural environments that take place in varied eras, made with Unreal Engine 5, and a charming soundtrack to make it a journey you may not soon forget. GENIE is a couple of hours of a relaxing experience. First Collect the broken crystal shards then follow the unraveling poems of the story. virtual) required for installing this repackĮxplore fantastic environments as Lula, as she journeys to discover the meaning of the Genie and fulfill her destiny. Use “GENIE (Windows 7)” desktop icon to play on Windows 7. ![]() Repack uses compression library by Razor12911.After-install integrity check so you could make sure that everything installed properly.Installation takes from 15 minutes (on 24-threads CPU) up to 1 hour 20 minutes (on 4-threads CPU).Smaller archive size (compressed from 24.2 to 18.5 GB).100% Lossless & MD5 Perfect: all files are identical to originals after installation.Based on GENIE-TENOKE ISO release: tenoke-genie.iso (25,991,794,688 bytes).Filehoster: OneDrive (Uploaded by DyR0 t(-_-t), compatible with torrent mirrors)ĭiscussion and (possible) future updates on CS.RIN.RU thread Screenshots (Click to enlarge).Filehoster: MultiUpload (10+ hosters, interchangeable).Genres/Tags: Exploration, First-person, 3D 7/23/2023 0 Comments Miches sip n dip![]() On any given night, you could hear everything from Sinatra-era crooners to outlaw country by Johnny Cash to lighthearted Jimmy Buffet’s “Margaritaville.” If you asked her, though, she might even play some of the more raunchy tunes she knew. If you requested it and she knew it, she'd play it. Inside Pat's head was a virtual jukebox full of songs. "She would always laugh and say, 'I don’t know why people always make a fuss over me,'” said Thares.Īnyone who's ever been to see her knows why, though. Thares caller her a gracious and humble person. We lost somebody that we loved very much.”ĭespite national and even global attention, Pat never considered herself a celebrity. ![]() "It’s a work family," Thares said. "We didn’t just lose a coworker. Thares and many of the Sip 'n Dip's staff had been together with Pat for 25 years. She also had a bad fall in 2020 that put her in intensive care, but Thares said she had recovered well. Sip 'n Dip owner Sandra Thares said Pat last performed in February 2020, but she was optimistic about being able to return to her piano. Rest In Peace beautiful music girl.” "We will absolutely 100% miss her"ĭuring the COVID-19 pandemic, Pat stayed home. “We loved her very, very much. Our hearts are broken. “It is with tremendously sad hearts that we tell you that our beloved Piano Pat passed away peacefully last night,” the Sip ‘n Dip wrote. On the Sip 'n Dip's Facebook page, more than 700 comments and climbing sit beneath the post announcing her death. Twitter immediately filled with dozens of posts from Montanans, ex-Montanans, politicos and journalists who came to know Pat and mourn her passing. The Sip 'n Dip Lounge will never be the same. Patricia "Piano Pat" Sponheim has played her last notes, passing away Tuesday in her mid-80s. "It’s just a quirky, weird kind of place.At a tiny tiki bar in Great Falls, generations of Montanans yelled "bum bum bum" to "Sweet Caroline" and "Where's the salt?" to "Margaritaville" crooned in a low, husky voice by a woman who spent almost 60 years as a beloved local legend. "We don’t try to be anything we’re not," says Thares. Sure, the fishbowls might double as bear tranquilizers - "I don’t know anyone who’s survived two of them," warns Mayor Kelly - but people are here to hang out over drinks, maybe eat some prime rib, and, of course, see the mermaid show. These all seem like elements of a rowdy roadhouse, but the Sip is surprisingly laid back. When they entered the bar for a cool-down drink, they received a standing ovation. And then there's the tale of the newlyweds who decided to consummate their union in the pool, unaware that the whole bar could see through the window. Mermaid Corbin's husband, a former merman, proposed to her underwater before he went back into active duty military service. ![]() The tank is also home to stories both heartwarming and salacious. "I put on the mermaid outfit and the little bow tie that Sandi gives people and I went in," Mayor Kelly told me. And during a local fundraiser about three years ago, Great Falls Mayor Bob Kelly became a merman. Legend has it, Splash star Daryl Hannah once donned a tail and hit the water while in town filming Northfork in 2002. You might even see a familiar face in the tank. That really ups the surreality factor when you enter the Sip ‘n Dip. Known as "Electric City" due to its stout collection of dams and power plants, this is a hard-working ranching and hunting community located next to an Air Force base specializing in ballistic missile maintenance. I know what you’re thinking, but Great Falls isn't some hipsterfied town where kitsch blends into the nightscape. On almost any given night after 9 o’clock, mermaids swim and perform behind the bar, often accompanied by the vocal stylings of octogenarian lounge singer "Piano" Pat Sponheim. This colorful tiki haunt is where Big Sky Country meets Polynesian basement vibes. The landlocked state is the last place you'd expect to see mermaids… but that’s exactly what you’ll find when you stop by the legendary Sip 'n Dip Lounge in Great Falls.īuilt in 1962 as part of the retro O’Haire Motor Inn, the Sip ‘n Dip just might be the best bar in America you’ve never heard of. Montana brings a lot of things to mind: dusty cowboys, grizzly bears, and outdoor adventure. 7/23/2023 0 Comments Silverback gorilla wallpaper![]() We are also inspired to learn more about conservation efforts and to take action in our own ways, whether that means participating in volunteer work or donating to conservation organizations. By looking at a gorilla wallpaper daily, we are reminded of the importance of the preservation of the environment and the protection of wildlife. Gorilla wallpapers serve as a tool to spread awareness, encourage engagement, and inspire action.Īnother reason to use gorilla wallpapers is that they inspire us to take action to protect these magnificent creatures. To protect these species, it is necessary to support organizations that work to protect habitats, stop illegal trade, and monitor and treat disease outbreaks. They are also vulnerable to disease outbreaks, which can rapidly spread through their communities. Gorillas are under severe threat from habitat loss due to deforestation, the bushmeat trade, and climate change. One of the primary reasons that gorilla wallpapers are so important is that they raise awareness about the conservation efforts needed to protect these animals. The gorilla has been used as a symbol for conservation, and it is often represented in artistic and creative ways. They are also of great importance to African culture and are very significant in traditional folklore. Gorillas are symbols of intelligence, strength, and friendship, which is why they are often featured in art and literature. Additionally, gorilla wallpapers serve as a testament to the importance of conservation efforts as the species is critically endangered. In many cases, gorilla wallpapers show the animals in their natural habitat, which brings a sense of peace and harmony to any room. Gorilla wallpapers are a fantastic way to pay tribute to these gentle giants and to remind ourselves of the beauty and complexity of the natural world. By featuring these incredible animals on your desktop or phone, you become part of the conservation efforts to protect this critically endangered species. Gorillas are majestic creatures that are on the brink of extinction due to habitat loss, poaching, and disease. You’re satisfied, press “set as wallpaper.” 8.If you’re a lover of nature and wildlife, then gorilla wallpapers are a perfect addition to your home or office to add a touch of beauty and inspiration to your everyday life. You’ll then be able to move the image to how you like. You’ll then be prompted to select whether you want to set the image as the background of Right corner, click the menu button (three vertical dots). The first image you see here should be the image you downloaded. Open your gallery/photos app and click on the “download” folder.ģ. Search for a wallpaper you like on and download it clicking on the blueĭownload button below the wallpaper. Navigate back to your home screen and take a look at your new wallpaper. Whether you want this image to be set as the background of your lock screen, home screen or both. Here you can arrange the picture how you want it, then tap “set.” 8. (the one that looks like a box with an arrow coming out of it). Navigate to the “Photos” app and find the image you want as your background. Tap on an image and hold on a few seconds. Find an image you like on and click on the blue download button Exit back to your desktop and see what it looks like! iPhone/iPad 1. Here you’ll want to select your own, so you’ll select the location your new imageĭownloaded. This order, click Apple Menu > System Preferences > Desktop & Screen Saver > Desktop 3. Download your favourite wallpaper clicking on the blue download button below the The folder and click “Set as desktop background.” 6.Enjoy your new wallpaper! Mac 1. That image on your computer (it will probably be in your “downloads” folder) 5.Right-click the image in That part for you.) 3.Click the button, and you’ll notice the image save to your browser. That says “Free Download.” Just below that text is your screen’s resolution (don’t worry, we calculated First, find the perfect wallpaper for your PC. 7/23/2023 0 Comments Docker mysql export databaserw- 1 systemd-coredump systemd-coredump 1680 Jul 1 11:07 server-key. MySQL, PostgreSQL, etc.) That’s pretty much it Download the latest version of Adminer from their official website. rw-r-r- 1 systemd-coredump systemd-coredump 1112 Jul 1 11:07 server-cert.pem How to Use Adminer But before you get started with installing it, here are some basic requirements to get Adminer working on your server: Install PHP 5, 7, or 8 A database driver (e.g. rw-r-r- 1 systemd-coredump systemd-coredump 452 Jul 1 11:07 public_key.pem rw- 1 systemd-coredump systemd-coredump 1680 Jul 1 11:07 private_key.pem rw-r- 1 systemd-coredump systemd-coredump 12582912 Jul 1 11:07 ibtmp1ĭrwxr-x- 2 systemd-coredump systemd-coredump 4096 Jul 1 11:07 mysqlĭrwxr-x- 2 systemd-coredump systemd-coredump 4096 Jul 1 11:07 performance_schema rw-r- 1 systemd-coredump systemd-coredump 79691776 Jul 1 11:07 ibdata1 rw-r- 1 systemd-coredump systemd-coredump 50331648 Jul 1 11:07 ib_logfile1 rw-r- 1 systemd-coredump systemd-coredump 50331648 Jul 1 11:07 ib_logfile0 rw-r- 1 systemd-coredump systemd-coredump 1346 Jul 1 11:07 ib_buffer_pool rw- 1 systemd-coredump systemd-coredump 1680 Jul 1 11:07 client-key.pem rw-r-r- 1 systemd-coredump systemd-coredump 1112 Jul 1 11:07 client-cert.pem rw-r-r- 1 systemd-coredump systemd-coredump 1112 Jul 1 11:07 ca.pem rw- 1 systemd-coredump systemd-coredump 1676 Jul 1 11:07 ca-key.pem rw-r- 1 systemd-coredump systemd-coredump 56 Jul 1 11:07 auto.cnf data/dbĭrwxr-x- 2 systemd-coredump systemd-coredump 4096 Jul 1 11:07 app_db ![]() To view these files, just run below command. In this case the MySQL container creats all files on host machine under. docker-compose.yml:Īfter creating file, just run the below command to launch container. Next create compose file in current directory. I run my export like this: docker run -it -link containername:mysql -rm mysql sh -c 'exec mysqldump -h'MYSQLPORT3306TCPADDR' -P'MYSQLPORT3306TCPPORT' -uroot -p'MYSQLENVMYSQLROOTPASSWORD' dbname' gz >. data/db as volume to store all files created by the MySQL server. I use the official MySQL docker image, and I am having difficulty exporting data from the instance without errors. Then configure docker-compose.yml to use. I am creating below directory structure under the current directory. In any case docker container get terminated, you can relaunch container using the existing data files.Ĭreate a directory to keep your MySQL data files. But, If you like, you can keep database files on the host machine. We recommend to use data volume instead of putting files on host machine. Option 2 – Storing MySQL Data on Host Machine You can view the docker volumes by running commnad: docker volume ls docker-compose up -dĬreating network "db_default" with the default driverĬreating volume "db_dbdata" with default driver Next, run below command to launch Docker container. This is the default directory used by MySQL to store all data files. The particular set of configuration options depends on the database type.The above configuration defined one data volume named “dbdata”, which is attached to MySQL container and mounted on /var/lib/mysql directory. You can pass a set of additional dump/restore parameters to the native tool. You can choose what schemas/tables you want to backup/dump: item and add a new client in the following dialog: ![]() To configure a new client location, choose the Browse. The native client configuration can be done in driver editor dialog or directly from the backup/restore wizard. The native client is a set of binaries (different for different OSes) which will be executed by DBeaver to process an actual backup/restore. In order to execute native backup/restore tools you need to configure the database native client. These functions can be accessed from the Context Menu's Tools or the Main Menu's Database->Tools. It uses database native dump formats and it may work much faster as it uses special utilities for the direct high-performance database access. The native backup restore differs from the standard DBeaver data transfer feature. Database Backup/restoreĭBeaver supports native database backup/restore functions for the following databases: NB: This feature is available in Enterprise, Ultimate and Team editions only. Importing CA certificates from your local Java into DBeaver.Connecting to Oracle Database using JDBC OCI driver.docker pull mysql Overview Tags Sort by Newest No tags found This repository doesnt have any tags that have 'mysq' in their names. Installing extensions - Themes, version control, etc Explore Official Images mysql mysql Docker Official Image 1B+ 10K+ MySQL is a widely used, open-source relational database management system (RDBMS). 7/23/2023 0 Comments Liquid quart to cups![]() That’s over 3 US cups more milk when you pay in pounds instead of dollars! So pay close attention to the origin of the recipe you’re using, since the author may be speaking a different language of measurement. But that difference becomes much more noticeable when you consider a gallon of milk, which in the US is 3,785 ml versus 4,546 ml in Britain. The difference in a teaspoon of vanilla would be hard to measure even if you tried. For example, a US contemporary teaspoon is 4.93 ml compared to 5 ml in the Britisth Imperial System teaspoon. These differences are small when the amounts are small, but can really add up for larger volumes. Even within the US, there are differences between the US contemporary system and that used by the US Food and Drug Administration. To add to the confusion, these systems all use the same names, such as pints and quarts, to mean slightly different measurement amounts. Some English-speaking countries, including the United States, United Kingdom, and Australia, use measurement systems that originated from an old system called “English units”. Most countries use the metric system (officially known as the International System of Units), where every unit is defined using a measurable phenomenon, such as the distance light travels in a second. For other substances, the density will be different, and each teaspoon will weigh a different number of grams. Water has a density of 1 g/ml, so the conversion is 1 gram to 1 millileter, which is equivalent to 0.2 teaspoons. The correct conversion depends on the density of the item you're measuring. ![]() Grams are a measure of mass, and teaspoons measure volume. If you're looking for a grams-to-teaspoons conversion chart, you won't find one here. 7/23/2023 0 Comments Never have i ever season 1 cast![]() There will be 10 episodes in the fourth and final season of the series. Mohan Vishwakumar, played by Sendhil Ramamurthy.Trent Harrison, played by Benjamin Norris.Paxton Hall-Yoshida is played by Darren Barnet.Nalini Vishwakumar is played by Poorna Jagannathan. Depicting Devi Vishwakumar as Maitreyi Ramakrishnan.The fourth and last season of "Never Have I Ever" will air on Netflix in June 2023. When is season 4 of "Never Have I Ever" scheduled? ![]() Thanks to Devi's educational programme in California, Paxton and Ben almost completely lost Devi, but she decided to stay since she wasn't yet ready to go home and say goodbye to her mother, Nalini.Īlso introduced is Ethan, a recent school darling who is sure to catch the eye of all the girls.ĭevi has a lot of choices to make, including which of the two males she wants to date, the direction of her schooling, and her interactions with her family and friends.ĪLSO READ: Never Have I Ever Season 3 Review: Maitreyi Ramakrishnan's series is crisp, witty and addicted to love However, despite Paxton's high school graduation and plans to attend a university in Arizona, there could be one more effort to rekindle the romance with Devi. What changes will Season 4 bring to the game?īen, Devi, and Paxton's love triangle continues as they start their final year of high school.ĭevi definitely let Ben know how she felt about him by showing up at his place to redeem his "one free beer" coupon. Here are certain details you should be aware of before the series airs, starting with its fourth episode. The series has received over a million hours of viewing time globally since Netflix began sharing its viewing data on a weekly basis for the global top ten rankings. Maitreyi Ramakrishnan as Devi VishwakumarRamona Young as Eleanor WongLee Rodriguez as Fabiola TorresDarren Barnet as Paxton Hall-YoshidaJaren Lewison as Ben GrossMegan Suri as Aneesa. The events from Kaling's own adolescence serve as a loose inspiration for the series. The coming-of-age comedy series Never Have I Ever was a Netflix original comedy that was produced by Mindy Kaling and Lang Fisher. 7/23/2023 0 Comments 10x chromium core mgh![]() A bird genome assembly showed a contig N50 of 147 kb and a scaffold N50 of 18 Mb for a 1.2 Gb genome. However, the Bioinformatics Core has generated and high quality genome assemblies with scaffolds comprising entire chromosome arms for both birds and mammals from our 10X data. ![]() Thus, the design is optimized for large genomes and works best for genome sizes around 3 Gb. Please note that the 10X Genomics Chromium technology was designed primarily for the analysis of human genomes. We offer the isolation of HMW-DNA samples – please see this page.ĭe novo Genome Assembly Projects: 10X Chromium sequencing data can yield excellent de novo genome assemblies when assembled with the free Supernova2 genome assembler. The 10X Genomics Chromium Genome assays do require paired-end 150 bp read sequencing. Thus nicks present in the DNA sample will significantly reduce the linked-read length. The first step of the actual 10X library prep is a denaturation of the sample. For DNA samples significantly longer than 50 kb these pre-treatments will likely not be necessary. For DNA samples at this size we might recommend two pre-treatments: DNA-Damage-Repair and BluePippin Sample Size Selection (the latter will remove fragments shorter than 40 kb). The DNA amount required for the actual library prep is minimal (1 ng), however more DNA is required for the QC of the sample (we suggest to submit at least 500 ng at a concentration of 20 ng/ul or higher as determined by Qubit).ġ0X Genomics recommends DNA sample should run as a band of 50 Kb length or higher on a pulsed-field gel. The quality of the DNA samples should be verified by pulsed-field gel electrophoresis. 10X Genomics does suggest the Qiagen MagAttract HMW DNA Kit for DNA isolations (for plants other DNA extraction methods will be required). The DNA quality will determine to a great extent the linkage information that can be gained from the 10X data – the longer the input DNA fragments the greater the length of the linkage information per droplet. The 10X Genome sequencing data contain conventional shotgun sequencing data, after trimming off the first 26 bases of the forward read. For human whole genome phasing and structural variant analysis GemCode sequencing data at 60x genome coverage or a combination of conventional Illumina data coverage at 30x genome coverage and GemCode at least 23x is recommended. Human phasing can be carried out using both whole genome shotgun sequencing as well as employing exome capture enrichment. A single HiSeq lane generates sufficient information for phasing and structural variant analysis of a human sized genome. Sub-genome sized parts of the DNA sample (about 1/6000th of a genome) are compartmentalized in nanoliter volume oil droplets together with beads carrying one of about two million different barcodes to initiate the library preparation with droplet specific barcodes. Please note that the individual DNA molecules are not meant to be fully assembled with this approach. DNA equivalent to about 150 copies of a genome does get distributed over about 1 million oil droplets that include beads. Thus, it allows individual long DNA molecules 10X Genomics Chromium Genome Linked-reads principle. The 10X Genomics technology generates individually barcoded sequencing libraries for hundreds of thousands of nanoliter volume oil droplets using up to 1.7 million different barcodes. Please see these slides for details on GemCode technology and some of the possible applications. Genotyping in repetitive regions of genomes is possible via an approach called “critical content rescue”. The applications currently supported by the 10X Genomics software are human haplotype phasing and structural variant detection (see this page for currently supported applications) as well as de novo genome assemblies (please see below). Just a few nanograms of sample ar e required as input for the library preparation. When analyzing high molecular weight DNA samples the linked read sequencing data delineate linkage information over distances of up to 150 kb. The sequencing libraries are generated with the Chromium controller instrument and reagents from 10X Genomics, followed by sequencing on our Illumina HiSeq sequencers. We are offering 10X Genomics Chromium Genome “linked read” library preps and sequencing. Please contact to reserve reagents for your project. As soon as these run out, we will discontinue the assay effective immediately. Update July 2020: We have ~16 remaining 10X Chromium Genome reactions left in our core. We are sad to see this very popular assay go. As of January 2020, 10X Genomics has announced that it is discontinuing production of the 10X Chromium Genome linked read reagents. |