Blunt ends are always compatible, so can always work together. Consider filling in an overhang: While not ideal, a short treatment with T4 DNA polymerase or Klenow can fill in a sticky overhang and make it blunt.Consider enzyme overhangs: Some enzymes have overhangs that are in common and can be used together, despite not being the same recognition sequence, for example, BamHI and BglII.There aren’t enzymes in common between the destination and insert: This is the most common issue you may run into.Some other common problems and workarounds: Since this is the common promoter used in my lab, I knew I could add it back in from another plasmid I had on hand. In my case, I noticed that while I couldn’t remove the guide sequences on their own, I could remove the guide and promoter. If this sort of thing happens to you, don’t worry! There are almost always creative solutions to these sorts of problems. As you can see, there aren’t any cut sites in between the old guide RNA and my promoter. This is where I started to run into problems. If you spend time carefully planning now, it will almost always pay off in the end.įor my particular cloning experiment I started by mapping the various enzyme sites on my vector. Try and make the process as simple as possible, so that the actual cloning is easy as well. If at all possible, use enzymes that only cut once within a vector, leave sticky overhangs (that is 5′ or 3′ DNA overhangs), and have simple recognition sites (usually 6-8 base pair sites). It is important to consider how many times the enzyme cuts in the vector, the overhang that is left behind, and the type of enzyme recognition site. For many cloning experiments, this can be complicated. I was only concerned with removing the old guide RNA sequences and replacing them with those that I had designed:įor restriction cloning to work, you need to find enzymes that cut both the target vector and the incoming DNA, so you need to examine the cut sites. The vector consists of the old guides, a plant ubiquitin promoter to express the Cas9, a selection gene, and various plasmid maintenance genes. The first file to look at is the destination vector, so I opened mine up: I use SnapGene, but there are a large variety of programs out there that have these capabilities. The first thing to do is open up your sequence files in your program of choice. Pull Up Your Target Vector, Desired Insert, and Other DNA Sequences on the Computer Screen I also knew the new guides were being synthesized and shipped to me in a standard vector, pUC19. I knew that I didn’t have to worry about anything being in frame, because that was already done for me. This seems simple, but it is the most important part of the planning process.įor my cloning experiment I needed to remove the guide RNA sequences in a Cas9 vector that I had, and replace them with a new set of sequences I had synthesized by a company. Make sure to make note of things like frame requirements, etc. Write down exactly which vector you have, what components you need where, and any other stipulations that are required at the end of the process. Lay Out What You Have and What You Need for Restriction Cloning It had a little of everything, so it will cover most of the problems you will run into in your cloning endeavors. We are here to help! The easiest way to learn anything is to do it, so I thought I would take you step-by-step through the last cloning set-up I did. If you are just starting with restriction cloning, it can be a bit of a struggle to learn the process, but that is ok. While getting each of the steps correct can be a bit of a hassle, in my mind, the most difficult part of the whole process is planning it. You simply cut the target vector and insert with the same enzymes, clean digested vector and insert up, ligate the two together, transform the ligated vector and insert into bacteria, and then screen. Restriction cloning, at its core, is quite simple.
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Check out your local tide charts in Ballena Tales for more details on the tides!įor first-time surfers, Punta Uvita/Playa Chaman has great waves to help you learn and get up on the board for the first time. The best tides for beginner surf lessons are the mid tides. You will find most of the surf camps in Dominical and Uvita. ![]() ![]() Where can you go for family surfing? The most commonly surfed beaches along Costa Ballena are Uvita, Dominical, Dominicalito, Hermosa, and Ventanas. Once you’ve had a few lessons and you feel confident, you can then think about renting a board and heading out on your own. The surfboards you are given are specially designed for beginner surfers, and depending on the camp, the instructor may take you to visit a few of the numerous local surf beaches. The instructors in the area are all seasoned surfers with great knowledge of the area. Not only are most surf camps cheap, they provide everything from boards, leashes, wax and sometimes a snack on the beach. For safety and fun, first-time surfers should be an instructor or an expert. Should families take surf lessons? It is not advised if you have never surfed, to rent boards for you and your family and just boldly rally to a nearby beach. Here are some tips to make your family surf adventure an experience that will make you come back for more. In Costa Ballena, Costa Rica, South Pacific with its numerous sandy beaches, you should never be far from a local surf spot. The equipment has gotten lighter, making it easier to maneuver, and both adults and children of all ages can enjoy taking lessons together. ![]() Thank you for reading this post, don't forget to subscribe!įamily surfing vacations have become increasingly popular in the last 10 years. ![]() Additionally, you can download videos off 1000+ websites, including Facebook, Dailymotion, Twitter, Instagram, and more with Y2mate 4K Video Downloader Free. The free 4K Video Downloader allows you to download videos in HD quality up to 1080p in seconds. The easy-to-use 4K Video Downloader free supports to download 4K video in 3 simple clicks with a simple interface. Y2mate is another best 4K Video Downloader free to download 4K video online. Instead of copy-pasting the URL of the video, this 4K Video Downloader free allows you to search for the YouTube videos directly on the website and download it to your device. VidPaw is a free 4K video downloader to download 4K videos online quite easily. Download all videos from websites online.Display the download speed and status of the complete video.Browse videos with the built-in browser.There is no need to register or log in to download your favorite videos with this 4K Video Downloader. This free 4K Video Downloader is totally free and downloads every video with a single tap. Doesn’t support downloading 4K video to Android phoneĪll 4K Video Downloader is a super-fast video downloader to download 4K video very easy way and fast.Supports iOS data transfer and file management.Download videos from BBC, Vimeo, Instagram, Facebook, etc.100% secure and free to download 4K video.The 4K Video Downloader free can transfer these videos to your computer by pasting the link/URL of YouTube video you'd like to download MobiMover is a professional free 4K Video Downloader that can not only help you efficiently manage iOS data but also download 4K video from Facebook, Twitter, YouTube, Vimeo, Dailymotion, and more. ![]() Equipped with a built-in video player to watch videos on your devices.Supports download 1080p videos from YouTube and other 50+ sites.The free 4K Video Downloader has a built-in video player. The 4K Video Downloader also supports downloading videos to different formats, such as MP4, MP3, WMV, FLV, MOV, 3GP, and more. YTD is another powerful 4K Video Downloader free for saving videos from YouTube and other 50+ sites. Part 2: 12 Alternatives to Free 4K Video Downloaders Recommended 1. Besides, we also recommend you Videoer and YouTube4KDownloader to download 4k videos and our YouTube to MP4 Converter to convert YouTube videos and save in high resolution.īelow, we have summarized 12 alternatives to 4K Video Downloader free recommended to download 4K videos. ![]()
A full path for a mailbox within an account such as A mailbox path without an account such as /Special (does not work for goToMailbox:).A standard mailbox name such as inbox, archive, drafts, sent, junk, and trash.The UUID of a smart mailbox (as saved in ~/Library/Application Support/MailMate/ist).A mailbox UUID is not always a UUID in a strict sense, but it is always a unique identifier for a mailbox. The goToMailbox: and moveToMailbox: selectors work both with and without an argument. The following table contains some of the most interesting examples. MailMate extends the list of selectors accepting an argument. ![]() "$\UF704" = ( "insertText:", "Thanks for trying out MailMate." ) This is a couple of the standard key bindings defined in macOS (converted to plain text property list format): A detailed description of key bindings has been written by Jacob Rus and this is recommended reading if you want to get the full story. The standard definitions are located in a file named in the so-called AppKit framework, but this file is in a binary format and viewing it either requires a property list editor as distributed with Apples developer tools or conversion using the plutil command line program. These key bindings affect most applications and most of them affect keyboard navigation in text fields or other views. The key stroke for quitting is fairly high-level in the sense that it is handled by each application individually and this is true for most key bindings involving the use of ⌘.īut there is also a more low-level system for key bindings in macOS. For example, the key stroke ⌘Q is most often bound to the action of quitting. BackgroundĪ key binding is the translation of a key stroke (or series of keystrokes) to some action. If you simply want key bindings similar to those available in the Gmail web interface then go to the General pane of the Preferences (⌘,) and enable the default custom key bindings. Bobbi Brown Skin Long Wear Foundation: Which is Best for You?īest Hair Oil Reviews: Kérastase Elixir Ultime vs. Shape Tape Foundation: Which is Best for You? (Review + 7% Cashback) ![]() YSL Wallet: Which is the Best to Invest in 2022? CHANEL Classic Camera: Which is the Best Designer Camera Bag to Buy Costa Del Mar: Which Makes the Best High-End Sunglasses Brand? ![]() Anjou Tote: Which is the Best Tote to Invest in 2022 (Sizes + Price) YSL All Hours: Which Foundation is Best for You? Gucci Bags: Which Brand Is The Best? (History, Design, Quality & Price) Artsy Review: Which of the Three Should Be Your First LV Bag?ĬHANEL vs. Tod's Shoes: Which Brand is the Best? (History, Quality, Design & Price)Ĭoach vs Tory Burch vs MCM Bag: Which Brand Is The Best? (History, Quality, Price & Design) Longchamp Tote: Which Will Stand the Test of Time?īally vs. Gucci Retail: Differences, Quality & Price 2022 Nike Retail Store: Differences, Quality & Price 2022 Retail Store: Differences, Quality & Price 2022 Retail: Differences, Quality & Price 2022 Emporio Armani Watches: Which is Best for Style?įossil Outlet vs. I'm sure you have found your answers! Sign up at Extrabux , then you can enjoy up to 5% cashback on your order! Shoppers can expect to find past seasons' merchandise and "affordable luxury" products manufactured specifically for the factory outlet stores. Value-conscious customers of Ann Taylor retail stores will find some of the same popular styles at discounted prices at LOFT Outlets and Ann Taylor Factory stores. However, LOFT Outlet and Ann Taylor Factory stores can help the retailer stay competitive. But with relatively fewer promotions in its full priced stores, Ann risks losing out to its competitors. However, LOFT Outlet and Ann Taylor Factory stores reach out to a larger customer base due to price discounts.Īnn's strong brand recognition and "affordable luxury" tag create a strong appeal. Most of Ann's merchandise fall under the "better" and "moderate" price category, implying that they are relatively expensive and cater to more affluent customers. So when a brand new item comes in that you have to have, but you don't want to pay full price, wait it out. Loft is always having sales, many that are 50% off select items. Also, there usually is a nice selection of petite sizes at both stores. Both divisions sell suits, separates, dresses, shoes, and accessories designed for professional women. LOFT carry a more youthful look and more casual styles, while the Ann Taylor stores cater more to professionals and business. While these items look like regular stores and have the same name, they are generally made from cheaper or less functional fabrics than their full-price counterparts.Īnn Taylor is a premier American specialty apparel retailer for the professional woman. Factory outlets sell products made specifically for outlet outlets. Outlet-specific merchandise is often of lower quality than goods sold at non-outlet retail locations and online store. Together, they encompass the entire range of Ann's product offerings. ![]() LOFT Outlet focuses on women's casual attire including casual separates, dresses, tops, weekend wear, shoes and accessories. There are many styles cheap than retail store of Ann Taylor and LOFT.Īnn Taylor Factory focuses exclusively on professional attire for women, offering them wear-now and wear-to-work fashion at affordable prices. ![]() Items in outlet and factory stores are usually up to 50% off. There are 139 Loft Outlets and 98 Ann Taylor Factory stores in the United States. Factory and Outlet store carry a mixture of last season's styles along with items made specifically for the outlet.Īdditionally, they offer products in the “affordable luxury” segment, designed specifically under the LOFT Outlet and Ann Taylor Factory labels. (7.5-9% cashback,second-hand items)Īnn Taylor Factory store and LOFT Outlet keep goods that are latest and they come directly from the warehouse, they are generally priced at the retail price unless there is some discount or something going on. Where to buy Ann Taylor and Loft the cheapest? sign up at Extrabux first! I tried on at AT recently and found the clothes to run smaller than Loft sizes.Ħ. I get the same size from both stores, but the Loft's fit is a little tighter on me than AT's. Are Ann Taylor and LOFT sizes the same? In my experience, the Loft's clothing in general runs a little smaller than Ann Taylor's. On one side of the stage stood two male percussionists - experts in performing palmas, traditional flamenco hand claps. They finished “Como Ali” - an unreleased tribute to Muhammad Ali - with a series of long, rapid-fire air punches, then held their hands overhead in fists. ![]() Counterpointing sexiness with a kind of SoulCycle strength, Rosalía’s troupe of female dancers wore white bike shorts and hoop earrings. Less visible was the garter-belt tattoo peeking out below her tiny, high-waisted shorts: a replica of the one that the Austrian feminist performance artist Valie Export gave herself onstage in 1970. Giant digital screens hung on either side of the stage, projecting her face to the crowd. But as she took the stage in white platform sneakers and an aqua top with an enormous ruffle running over her arms and chest, she gushed in Spanish, “I am so, so, so thrilled to be here!” For her, Mad Cool was a quick stop in the middle of a nine-month tour through Latin America, Europe and North America. Wednesday, however, was for Rosalía, Spain’s greatest pop export since Julio Iglesias. Heza Sport, Laver and The Falcon were all great first up but didn’t run on as well as expected on Friday night.One Wednesday in July, 40,000 people gathered on a synthetic-grass field on the outskirts of Madrid to watch Rosalía headline the Mad Cool festival’s “welcome party.” For the next three days, 103 acts - including Bon Iver, Iggy Pop and Prophets of Rage - would appear at the festival grounds, which were ornamented by a towering white Ferris wheel. There were several head-scratching efforts in the New Brighton Cup mainly from the horses that filled the placings in the Maurice Holmes Vase. Smiffy’s Terror ran on strongly to take third behind the Stonewall pair. “He is safe away but not fast, hopefully over the next six weeks we can get back in the draw and get him a little bit quicker away so he doesn’t have to work quite so hard when he is racing the absolute best ones.” “The only thing we have got to work on with him is the start.” The four-year-old was last at the 400m but charged down the outer, passing all but one of his rivals in the home straight.ī D Joe ran his last mile in 1.54.55, almost a full second faster than any other horse in the New Brighton Cup. “Most times he uses his manners and speed to win, but it is definitely good to see him strengthen up a bit.”ī D Joe was outstanding in second behind his stablemate. “He did lead and win the sales race at Kaikoura last year, but that wasn’t in this class.” “I think it is a progression that he has got stronger and is now able to do what he did tonight.” ![]() “The thing we pride him on is his manners, he can step and take a trail,” Telfer said. “He looked like almost anticipated the start and got himself into a bit of a bad spot, but he regained himself quickly which was good.”Īlta Wiseguy’s speed has never been questioned and it was on full display during his wonderful spring and summer campaign last year.īut in his returning win on Friday night, it was the horse’s strength that shone through.Īlta Wiseguy was pressured before the bell by Laver, but kicking up to hold the lead didn’t take a toll on the horse as he powered to victory. Steve Telfer thought it might be a case of Alta Wiseguy being a little too clever for his own good. The only concern for Alta Wiseguy fans came immediately after the start of the New Brighton Cup, with the four-year-old galloping away.īut in a matter of seconds, the pacer was back in his gear and making the kind of brilliant beginning he is known for. Show Me Heaven then made a huge night even bigger with her late closing win in race 10. ![]() The quinella was just one part of a brilliant night out for the Steve and Amanda Telfer stable with Aardie’s Express just as brilliant in her Nevele R Fillies heat win and Alta Meteor crushing from the death seat in his two-year-old assignment. Trying to be a little too wise wasn’t enough to stop Alta Wiseguy from booking his ticket into the New Zealand Cup on an incredible night out at Addington for Stonewall Stud.Īlta Wiseguy led home his stablemate B D Joe in the New Brighton Cup with both horses producing most outstanding lead-up efforts in the countdown to the country’s greatest race. Restart DSpace and then type the following to test. If the test build works then add the switch"-Pxpdf-mediafilter-support" to the dspace rebuild script, so that: To install, type the following: (Replace XXX with your DSpace version number)Ĭd $HOME/source/dspace/target/dspace-XXX-build Mvn -U clean package -Pxpdf-mediafilter-support Then replace ". = Adobe PDF" with the following:į. = Adobe PDFį. = Adobe PDFĪbove the comment, "#Custom settings for PDFFilter", add the following: BrandedPreviewJPEGFilter = Branded Preview JPEG PowerPointFilter = PowerPoint Text Extractor, \ ![]() Search for "ugins" and replace with the following.Ĭhange the MediaFilter plugin configuration to remove the old ".PDFFilter" and add the new filters ".XPDF2Text = PDF Text Extractor" and ".XPDF2Thumbnail = PDF Thumbnail". # maximum width and height of generated thumbnails Lastly, be sure there is a value for thumbnail.maxwidth and that it corresponds to the size you want for preview images for the UI. Then, search for the following and change as needed: Nano $HOME/source/dspace/config/dspace.cfgįirst enable thumbnails, search for following and change to true: Wget -no-check-certificate mvn install:install-file \ Step 3B - Install "jai_core.jar" mkdir $HOME/temp Dfile=jai_imageio-1_1/lib/jai_imageio.jar \ ![]() Step 3 - Install the java packages Step 3A - Install "jai_imageio.jar" mkdir $HOME/tempĬurl -O tar -xzvf jai_imageio-1_ Sudo apt-get install xpdf poppler-utils curl This filter has been disabled as these branded previews are not important to us.Ĭomplete ALL of the following as the "dspace" user! Step 2 - Install the Ubuntu software packages We isolated the problem to the "Branded Preview JPEG" filter. 12 Step 10 - Item list preview settingsĪfter a while we noticed our server load increasing radically when doing the nightly media-filter jobs.8 Step 6 - Update dspace rebuild script.5.1 Step 3A - Install "jai_imageio.jar".4 Step 2 - Install the Ubuntu software packages. ![]() It’s abstract gameplay and bizarre themes give it an air of mystery that is often unseen in modern dystopian science fiction. There’s no save function, so this is something designed to be played in one sitting. Expect a heavy narrative experience with this one that does away with perplexing puzzles and awkward combat in favour of eager discovery and exploration. Most recently he is known for playing one of the scientist guys in Guillermo del Toro’s The Shape Of Water. The Alpha Device is a surreal first-person adventure game narrated by David Hewlett no less. It’s that claustrophobic atmosphere that really sets Beherit apart as a truly unnerving game, and why you should definitely play it. ![]() This short but intense horror experience teases you with the idea of what could be lurking in the shadows rather than straight up show you. Trapped alone in this nightmare, it’s your task to uncover the hidden truth behind the mysterious gemstone known as the Beherit to give you any chance of escape. It’s not only the constant creaks that’ll keep you on edge though – there’s also the sporadic cracks of lightning to contend, with as if the horrors stalking the hallways weren’t enough. This first-person, horror exploration game thrusts you into some kind of creepy guest house where every door hinge is apparently badly maintained. Beheritīuckle up, because a lot of games on this list are in the horror genre and Beherit is no exception. So if you like horror and don’t have a lot of time on your hands this short, scary tale might be just the right fit for all you budding detectives out there. This point and click adventure has multiple endings and a bunch of hidden details for you to discover. Sat at your computer, you’ll read notes, decipher codes and unravel the unnerving truth behind a slew of grotesque murders. In this short horror adventure, you’ll find yourself in what looks like a mid-90s office hellbent on uncovering the mystery that lies within a box of strange police case files. Who doesn’t love a good murder mystery? Nobody, that’s who, and Location Withheld is exactly that, among other things. So, let’s dive right into the 100 best free indie games of all time. Whether they’re just starting out or because they want their story heard, there’s always going to be something interesting being said. Firstly because they’re, well, free – but also because developers who put time and effort into free games are doing so because of their passion. Many free games are just as good as, or even better than paid games. ![]() The idea that free games must be low quality is a common misconception. ![]() Choosing a Covenant is the most important decision that your character will make in Shadowlands, as the Covenant abilities and Soulbinds will significantly impact your spec's gameplay and performance. Shadowlands introduces several new expansion-specific features to help customize your character in the form of Covenants and Legendaries. The best WeakAuras to track the most important aspects of your gameplay. The best Macros and Addons for your Restoration Shaman How to optimize your rotation and increase your performance in every situation. ![]() Learn more about the best talent builds for Restoration Shaman. Your one-stop-shop for all the important information about how to play Restoration Shaman Looking to start playing your Restoration Shaman? Our starting guide provide all the information you're looking for, including the best talent builds, how to execute the proper rotation, and the best companion tools for your class with Macros, Addons, and Weak Auras. Restoration Shaman Starter GuideSeason 4 Guide for Restoration Shamanįor more information on how to play your Restoration Shaman, please see our Rotation Guide or Talent Guide. We also have an in-depth guide to Season 4, highlighting all the important aspects for the experimental season. If you are looking a brief overview of how to play a Restoration Shaman in Shadowlands, please visit our Starter Guide for a short summary on Covenants, Legendaries, Stats and more. Restoration Shaman in Shadowlands Season 4 When we do have to move though, we can shift into Ghost Wolf and cover a lot of ground quickly, but are unable to cast spells while transformed. Restoration Shaman aren't very mobile and love to just plant on a spot, spamming healing spells, especially because of our long cast times. This is accompanied by strong cooldowns which can get you out of dangerous situations easily, quite a bit of utility, and the ability to restore Mana to ourselves and allies. We have some supportive Healing over Time spells, and decent single target heals. ![]() We have a wide range of spells and tools to do our job, most of them being more effective when the group stacks closely together. We can see far into the distance with Far Sight and have our own separate Hearthstone with Astral Recall. Restoration Shaman call upon the healing properties of water and their iconic totems to keep their group members alive using Mana as the primary resource. How to Play Restoration Shaman in Shadowlands Restoration Shaman Strengths and Weaknesses For questions you can find me on the aforementioned discord servers or during my mute streams on Twitch. Also responsible for the Restoration Shaman sections of WoWAnalyzer and contributed towards Simulationcraft to make simming Restoration Shaman DPS possible. This page is maintained by Niseko, Restoration Shaman for CyaThursday on Stormscale-EU, co-creator, author and theorycrafter for Ancestral Guidance, admin in its Restoration Shaman specific Discord Server and Moderator in the Shaman Class Discord. Make sure to navigate to additional pages of the Restoration Shaman guide to find more in-depth information on topics such as Covenants, Legendaries, Soulbinds, Conduits, Tier Sets, Mythic+, and Raids.Ĭlick here for the Classic Version of the Restoration Shaman Guide! Throughout this guide, we will cover many different aspects to increase your Restoration Shaman expertise, including concepts like Restoration Shaman talents and talents builds, Restoration Shaman BiS gear choices, Restoration Shaman stat priorities, among many other aspects of your class and specialization. Welcome to Wowhead's Restoration Shaman guide, updated for ! This guide will teach you to master playing Restoration Shaman in all aspects of the game, helping you to keep your allies alive in Raids and Mythic+ dungeons. ![]() Launch the GoodNotes app in BlueStacks and create an account.Search and install GoodNotes from your Play Store, just like any android app.Start the BlueStacks app player on your PC, and go to its Google Play Store.Download and install BlueStacks on your Windows PC. BlueStacks is available on the google chrome app store.Follow the following steps to install GoodNotes Windows using Bluestacks. If you’re uncomfortable using VirtualBox, it is the more straightforward way to use android applications on your computer. With BlueStacks, you can run all Android apps on your computer. Download GoodNotes for Windows using BlueStacksĭownload & Install GoodNotes For Windows Using BlueStacksĪnother popular way to run GoodNotes on Windows is through the BlueStacks Android emulator.Install GoodNotes 5 from the virtual machine on the PC.Start running the virtual machine and mount the GoodNotes.Execute codes flawlessly to create a new virtual machine in VirtualBox.Download 64-bit VirtualBox on Windows PC and install it.Here are the simple steps to download GoodNotes for PC. You can download a virtual machine using VirtualBox. You’ll need to download an emulator to load the macOS on the virtual machine of your Windows system. Download & Install GoodNotes For Windows Using VirtualBoxĪ direct download of GoodNotes 5 for windows is unavailable in the Chrome Webstore. There are two popular ways to install GoodNotes on Windows PC. While talking about PCs, you can’t install it directly on the PC. Most people have started using this application regularly due to its versatility. ![]() With the rise of digital technology, GoodNotes has replaced traditional paper documents. The GoodNotes app for windows is the best option if you want to take your work anywhere. You can save your digital papers in the library and access them anytime. The template categories include homework, meetings, and letters to help you quickly enter any notes. With hundreds of pre-made templates available, this application is free for Windows PC. Using GoodNotes, you can take quick notes and organize your documents digitally. It is a customizable software that allows you to transfer notes from your iPhone, iPad, and iCloud to android. GoodNotes is an application to read and write PDF files. Download & Install GoodNotes For Windows Using BlueStacks.Download & Install GoodNotes For Windows Using VirtualBox. |
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