If you prefer to write using your mouse, then Avro gives you on-screen keyboards.īoth Bangla and English characters are available in the same fonts. Assamese characters are also possible with all of the keyboard layout options. You can type ‘Reph’ at the end of consonant characters and Avro will automatically move the ‘Reph’ to the beginning of the consonant. If you type a short version of a vowel, then the long-form will automatically appear. ![]() ![]() The keyboard layouts let you type easier and quicker with the automatic vowel formation feature. You can add and edit words within the auto-correct directory. The built-in dictionary lets you completely customize the autocorrect. Avro delivers an in-app dictionary with 150,000 Bengali words. A floating preview window will be beneath the letters to let you see the translations in real time. You can write Bengali words with Roman numerals to automatically translate the text to Bangla characters with the phonetic feature. How do you type on Avro Keyboard?Īvro Keyboard delivers phonetic-, touch-, and mouse-based typing. Avro Keyboard will track both the Bangla and default keyboard modes throughout all the applications on your PC for a seamless user experience. Additionally, the CTRL and spacebar keys will let you change the mode. You can select a hotkey from F1 to F12 that will quickly let you switch between keyboards. The current language will be shown in the toolbar. You can simply press the interactive button to automatically change the characters. The second icon in the top bar lets you open a dropdown menu to change the keyboard from the English language to Bangla. If you click on the Avro image, then you can switch the app back to the toolbar view: ‘ Restore Avro Top Bar’. You can click on the upward-facing arrow to open Avro Keyboard in the system tray. If you prefer to have Avro Keyboard in the system tray instead of the top bar, then you can tap on the logo to select ‘ Jump to system tray’. Additionally, you can move the top bar around the screen. Avro Keyboard is a highly customizable program that lets you make the toolbar transparent. The National keyboard layout is the standard format in Bangladesh. While OmicronLab released the ‘Avro Easy’ virtual keyboard layout to simplify typing, the Bornona keyboard is known to be the easiest Bangla layout to use. Additionally, you can press the ‘Show active keyboard layout’ button at the bottom of the expanded menu. The active layout will be marked with a bullet point. The keyboard layouts that are available are ‘Avro Phonetic (English to Bangla)’, ‘Avro Easy’, ‘Bornona’, ‘Munir Optima’, ‘National (Jatiya)’, and ‘Probhat’. You can hover above the ‘Select keyboard layout’ choice to expand the context menu. If you click on the logo button, then you can get a variety of options: ‘Avro Mouse’, ‘Spell checker’, etc. The top bar will display icons at the top of your desktop screen. How do I change my Avro Keyboard layout?Īvro Keyboard has a GUI with two main modes: top bar and system tray. While ANSI is optimal for certain applications, the Unicode mode is recommended for most circumstances. You can switch between the outputs by clicking on the settings icon within the top bar. Since Avro does not contain in-app advertisements, you will not have to worry about tracking by invasive spyware while using the tool.Īvro Keyboard is ANSI and Unicode compliant. While the original Avro Keyboard application is secure, variations of the open-source app may contain malware. Developed by OmicronLab, the utility gives Bengali speakers the opportunity to express themselves online. Title availability is subject to change.Avro Keyboard is completely safe to download and install. Some features may not be available in all countries or all areas. ![]()
0 Comments
It is one of the most intriguing places to see in Kuching, Sarawak. ![]() Believed to be over 100-years old, the Longhouse has been greatly expanded with interconnected walkways forming a proper community village. The government has also identified several locations in Semporna, Lahad Datu, Kudat and Kunak where stateless people may be relocated for better monitoring and management.Īrmizan said the centre could also potentially become an educational and arts centre that could attract tourists to learn about the Pala'u culture. 8 hours ago &0183 &32 Datuk Armizan Ali (pic) said this move is to help determine the exact size of Sabahs foreigner population. The Bidayuh Tribe, who make up 8 of Sarawak’s population, are the key residents at the Annah Rias Longhouse. “They will be sent to a temporary shelter for isolation for a maximum period of three months,” the minister said. “This centre offers a special programme that applies skills in self-management, community life and informal spiritual learning to beggars and detained street children. The population of East Malaysia is ethnographically even more complex than that of Peninsular Malaysia. It is the third largest island in the world, surpassed in size by only Greenland and New Guinea. ![]() He added that the majority of those sent to the centre were Pala’u, a group traditionally living in the coastal regions and islands of east coast Sabah.ĪLSO READ: Sarawak issues temporary papers to 86 stateless children Borneo, island in the extreme southwestern part of the Pacific Ocean. (decapoda: sergestidae) from the coastal waters of bintulu, sarawak: population structure. “By obtaining complete information about foreigners through digital data including facial recognition, it will enable the government to make a clear policy, including strategy and enforcement action mechanisms,” he said in a parliamentary written reply to Datuk Andi Muhammad Suryady (BN-Kalabakan) on Friday (May 26).Īndi had asked about holistic and integrated mechanisms by the government for managing issues related to the status of stateless persons, especially on Sabah's east coast.ĪLSO READ: Sabah testing out digital systems for special foreigners cardĪrmizan, who is Minister in the Prime Minister’s Department (Sabah, Sarawak Affairs and Special Functions), said the state government has also established a Non-Citizen Isolation Centre for beggars in Kota Kinabalu. The Distribution and Population Density of Bornean Tarsier. ![]() PETALING JAYA: The Sabah government is collecting biometric data to keep track of non-citizens, especially stateless persons, in the state.ĭatuk Armizan Ali ( pic) said this move is to help determine the exact size of Sabah's foreigner population.ĪLSO READ: Set up special committee to tackle street children issue, Sabah govt urged ![]() This article teaches you how to backup your iPhone, iPad, and iPod Touch using iCloud or iTunes. The next step is to download and install iTunes to reinstall the iOS. When a Live Photo appears on the Lock Screen, press the magnifying glass to view it. It will not be able to be played on your home screen because Live Photo is incompatible with that. If you don’t have an iOS version, tap Live, then choose a Live Photo you’d like to use. Live Photos can be wallpapered on iPhone 6s, iPhone 6s Plus, and later. Before proceeding, you must first make a backup of your data. Following that, you’ll need to reinstall the iOS using iTunes. If you have any questions or want to double-check the steps, please let us know. ![]() After a while, you’ll be able to start the conversation again by asking a new question. It appears that a live photo cannot be animated on the lock screen. ![]() SE, 5s, 6, 6 Plus, SE (2nd generation), iPad, iPod, and other devices are not supported. Other iPhone, iPad, iPod, and Apple TV models are not capable of displaying Live wallpaper on the lock screen only devices with the most recent version of the 6S can do so. The live wallpaper feature requires you to use 3D touch. Why Won’t My Live Photo Play On My Lock Screen? Live wallpapers can be installed on a number of Android devices, including the Google Pixel and Pixel XL, the Samsung Galaxy S8 and S8+, the LG G6, the HTC U11, and the Huawei Mate 10 Pro. If you’re just learning to use live wallpapers, we recommend looking through our list of the best free iPhone live wallpapers. Live wallpapers come in a variety of shapes and sizes, so you should thoroughly investigate each one. If you want something special to add to your phone’s personality or if you want to show off to friends and family, a live wallpaper is an excellent choice. Some of them are simple, such as a plain white background with a few small objects in the center, while others are more elaborate. Live wallpapers can be downloaded for free. The Stalker Cat, in particular, has a lovely blue and black gradient background, as well as a cute little kitten that welcomes you to its den. It has a plethora of beautiful fish, wonderful water rippling effects, and a plethora of nature scenes. Water Garden Live Wallpaper is a good choice for those looking for a water-themed wallpaper. Although the software does not require a PC to work, it can be used on an Android phone as well. The Wallpaper Engine app is a companion app for the popular Steam Wallpaper engine. The Walloop app allows you to create your own live wallpapers. The most popular of these are Paperland, Light Grid, and Minima. Live wallpapers work well, don’t consume a lot of battery, and have the right mix of activities without getting too worked up. Joko Interactive, a Google Play developer, has built a number of live wallpaper apps. You can find them in the Google Play Store. ![]() This content is written, edited and updated by hand surgeon members of the American Society for Surgery of the Hand. © 2014 American Society for Surgery of the Hand The sterile dressing will need to be changed two or three times a day, depending on the severity of the burn. Your hand surgeon will coordinate with a therapist if he or she feels that you would benefit from rehabilitation. A 2nd-degree burn that affects less than 10 of the skin's surface can usually be treated on an outpatient basis using antibiotic ointments. Deeper ones or those that occur in conjunction with other injuries may require extensive therapy and rehabilitation. Superficial burns generally will not need any formal hand therapy. Long-term follow up with your hand surgeon should be planned to evaluate for potential future surgery that could improve your hand function. Many burned hands will develop contractures, which cause stiff and constricted muscles, months or years after the original injury. ![]() Many patients with severe injuries will require more than one operation. Ideally this is within two to three weeks after the injury. After surgery, the hand is splinted (Figure 3) until the skin is healed. ![]() The dead skin will need to be removed and replaced with skin grafts, which is a surgical procedure. Wound healing occurs within two to three weeks. If there are open and raw surfaces, cleaning, frequent dressing changes, and the application of local topical antibiotics should be performed until the wounds are healed. The hand and forearm should be splinted in a position of safety to prevent later stiffness. They usually heal within a week.īlisters on the hand may or may not be trimmed. These are treated primarily for comfort with local pain killers. Those just on the hand may be treated on an outpatient basis if the pain can be adequately controlled. Burns over a major percentage of the body require hospitalization special care. How to treat a burn depends on the severity of the burn. Treatment focuses on preventing further problems with stiffness and infection. Initial first aid measures include removing the hand from the source of heat and keeping it clean. ![]() Archived October 29, 2007, at the Wayback Machine ^ Mark Chediak Suppliers of organic milk draw criticism.Dairy Reporter/Food Navigator, 18 August 2006. ^ Lorraine Heller Horizon dairy targeted for lacking organic practices."Danone CEO: Here's Why We're Buying WhiteWave". ^ "Horizon Celebrates 20 Years of Organic Leadership".^ "Happy Birthday Horizon Organic!" (PDF).^ "Dean Foods Company Completes Acquisition of Horizon Organic Holding Corporation".The acquisition was completed in April 2017 and newly formed company is named DanoneWave References In July 2016, it was announced that the French company Danone would purchase WhiteWave Foods for $10.4 billion. In 2013, WhiteWave Foods (owner of the Horizon brand) became an independent company from Dean Foods. In 2010, Horizon published "Organic Standards of Care" guidelines which outlines its livestock and organic dairy practices In 2009, (now former) parent company Dean Foods was sued by a group of dairymen for allegedly creating a monopoly on milk in the Southeastern U.S. As a result of the boycott, one of the largest natural food cooperatives, PCC Natural Markets in Seattle, decided to discontinue Horizon Organic products from their customer offerings. In August, the Cornucopia Institute filed a complaint with the USDA alleging that Horizon Organic violated National Organic Program standards in reference to the amount of time their dairy cows should spend in pasture. In 2006, the Organic Consumers Association organized a boycott of Horizon Organic (and Aurora Organic Dairy), alleging that the company was not following organic standards. The company went public in 1998 and was acquired by Dean Foods in 2004. The company sponsored a local K-8 school's extracurricular activities and even named their mascot, the Horizon Heifers. Horizon was founded in Boulder, Colorado in 1991 and began selling nationally in 1994. ![]() ![]() Horizon Organic's products are certified organic by the USDA and advertised as free of antibiotics, growth hormones and pesticides. Prior to being acquired by (now former parent company) Dean Foods, Horizon was a public company and owner of the organic dairy brands: Juniper Valley Farms, The Organic Cow of Vermont and United Kingdom brand Rachel's Organic. Horizon produces organic milk in Maryland and also purchases the majority of its organic milk (more than 99 percent) from more than 700 dairies in 23 states. Its products are sold in supermarkets and grocery stores. Horizon Organic is the largest supplier of organic milk in North America. Horizon Organic is an American company, founded in 1991, that produces organic milk and other organic food products. Prizes of up to $2,500 can be paid out on the spot by an authorized Pennsylvania Lottery retailer. How long does it take to receive PA Lottery winnings? This article originally appeared on : Powerball winning numbers drawing for Saturday, Feb.You can find the latest PA Lottery jackpot amounts for all of your favorite games here on the Pennsylvania Lottery Jackpots page. $536 million, Mega Millions July 8, 20116: Won in Indiana $543 million, Mega Millions, July 24, 2018: Won in California 11, 2015: Three winners in North Carolina, Puerto Rico, Texas 28, 2012: Two winner in Arizona, Missouri $590.5 million, Powerball, May 18, 2013: Won in Florida 5, 2022: Two winner in California, Wisconsin 17, 2013: Two winners in California, Georgia $656 million, Mega Millions, March 30, 2012: Three winners in Illinois, Kansas, Maryland $768.4 million, Powerball, March 27, 2019: Won in Wisconsin $1.337 billion, Mega Millions, July 29, 2022: Illinois 13, 2016: Three winners in California, Florida, Tennessee Here's a look at the top jackpots were won in the United States, between the Powerball and the Mega Millions lotteries: $768.4 million, March 27, 2019: Won in Wisconsin Here are the Top 10 jackpots since the Powerball lottery began in 1992: Check with your state lottery for more information. In other states, a winner's name and hometown are a matter of public record. That wasn't always the case, but now winners are able to stay anonymous under a law that was signed by Gov. In some states, like New Jersey, you can win a lottery anonymously. You don't have to wait decades for all the money. The cash option is significantly lower than the advertised jackpot, but it is paid in a lump sum. There is an immediate payment and then 29 annual payments after that, increasing by 5% each year. The annuity option is paid out over time. ![]() ![]() The major lotteries in the United States offer two jackpot payout options: annuity and cash. Unlucky? Here are 13 crazy things more likely to happen than winning the lottery Lump sum or annuity? The odds to match all five white balls are 11,688,053-to-1. The odds of winning the jackpot are 292,201,338-to-1. Playing the Powerball can be exciting, but just don't go spending those millions before you win. The drawing may be broadcast on a local television station in your market as well. You can watch the drawing by clicking here. The Powerball drawing is broadcast live on the lottery website at 10:59 p.m. Powerball, Mega Millions: These are the luckiest states for jackpot winners How can I watch Powerball drawing? Here is where you can buy tickets from lottery retailers in New Jersey: Click here. citizen or a resident a particular state where you purchase your ticket. Check with your state lottery for more details about online sales before purchasing tickets online. Some states allow Powerball tickets to be purchased online, but beware of scam websites. Many grocery stores, gas stations and convenience stores sell lottery tickets. You can play the game in 45 states plus the Washington DC, Puerto Rico and the U.S. New Jersey: Online lottery tickets sold by NJ could be an option for players next year Where can I buy Powerball tickets? You can check all the prize payouts on the Powerball website here. Prizes vary from $4 for the matching the Powerball to $1 million for matching all five white balls (except in California) to the jackpot for matching all six balls. However, you can also have the lottery machine generate a quick pick ticket with random numbers for you. There is also a 10x Power Play possibility when the jackpot is less than $150 million.Įach player selects five numbers from 1 to 69 for the white balls and one number from 1 to 26 for the red Powerball. The cost is $2 per ticket, but you can add the Power Play for $1, which will increase the amount of your potential prize up to five times the original prize (except for the jackpot and Match 5). on the day of the drawing, while New York's deadline is 10 p.m. The deadline for purchasing Powerball ticket varies by state so don't wait until the last minute. Powerball drawings are held three times a week - Monday, Wednesday and Saturday at 10:59 p.m. One ticket sold in Pennsylvania matched all five white balls to win $1 million. No one won it all so the jackpot for Monday will be $45 million with a cash option of $23.7 million. How to spot a scammer Did anyone win last night's Powerball? ![]() Beware: No, a lottery jackpot winner isn't giving you money. Mike tells the story of how it all came to be. The Fly Shop in Redding California is one of the largest online. Mike Mercer's Rod and Fly Shop was the first shop he created in his bedroom. We talk about the key to catching fish consistency, how to make a yarn indicator, the importance of the riffle drop and why you should avoid a tapered leader.ĭeli Fresh Design: use coupon wfs20 for 20% off) Mike shares the step x step plan on catching fish on the lower Sacramento. We hear how the Sacramento went from a toxic superfund dump with very few fish to a big time trout river. Various arts and crafts projects are also featured.I chat with Mike Mercer about the Sacramento River, the Redding Fly Shop and other great rivers around Chico California. With a display of lights, Christmas trees and the real Santa and his Elves this is a must see event! Stop in and get your child’s photograph with Santa. ![]() The highlight of the year is Santa’s Wonderland that is truly a sight to behold! During the holiday season, beginning in mid-November, the Tracker Boats area of our store is magically transformed into a breathtaking Christmas Wonderland. For service on all makes and models check out our Power Pros department featuring a state of the art three bay service center.īass Pro Shops features numerous events throughout the year such as the Spring Fishing Classic, Fall Hunting Classic, Go Outdoors, Photos with the Easter Bunny, and Family Summer Camp. In addition we offer a great selection of ATVs and UTVs. Our store features a complete selection of boats and motors, and a staff of experts to make sure you get the right combination. Tracker Marine is the world’s largest manufacturer of fishing boats. We are a pet friendly store so please bring your four legged best friends with you on your next visit! Need your bow worked on? Our associates will be happy to assist you. ![]() If you are interested in trying one of our bows before purchase, you may do so! The range is open from 9AM – 9PM Monday – Saturday, and 9AM – 7PM on Sunday. No crossbows please and only field tip arrows in the range. You can practice your shot in our 20 yard long archery range. ![]() The best part is that every department is staffed by welcoming knowledgeable associates who are eager to assist you!Ĭome get your bow sighted and ready for the hunt in our archery department. ![]() In addition to full Hunting and Fishing departments we feature Camping (including hiking, grilling, and everything in between!), Apparel and Footwear (with brands like Red Head, Under Armour, Columbia, Bob Timberlake and more), Gifts and Nature Shop (home décor, gifts, music, etc), Marine (boat electronics, accessories, GPS, etc), Water Sports (kayaks, rafts, and other fun water sports toys), Tracker Boats (fresh and salt water boats, big and small for fishing, hunting and fun. Northern California offers myriad outdoor lifestyle opportunities and Bass Pro stocks products to enhance them. We have weekly fish feedings on Saturday and Sunday at 1 PM. Extensive displays of indigenous wildlife, wall to wall murals of local landscapes, hundreds of antiques, and a 12,000 gallon aquarium combined with our complete collection of products make our store a regional destination to shop, browse and enjoy!Įnjoy viewing our 12,000 gallon freshwater aquarium that is stocked with fish native to our Northern California area including largemouth bass, striped bass, catfish, smallmouth bass, and sturgeon. Situated at the doorway to the Sierra, Bass Pro Rocklin features the area’s most complete selection of outdoor gear, apparel, and accessories embracing Northern California’s active lifestyle. Our 100,000 square foot store is conveniently located in the Cross Roads Shopping Center at I-80 and Sierra College Boulevard. Its fast trigger and low glow infrared will freeze the action day or night without spooking wildlife. Buy in monthly payments with Affirm on orders over 50. Start seeing the action as it’s happening even when you’re not in the woods Display: 24 per page. Tactacam Reveal X Tactacams Reveal X Cellular Camera is the ultimate remote scouting tool. ![]() Start seeing the action as its happening even when youre not in the woods The REVEAL Cellular Camera is not only small and discreet, but powerful enough to. The camera is ideal for locations with limited access or to put on your hunting properties. ![]() The camera is ideal for locations with limited access or to put on your hunting properties. Tactacam Reveal X VZN Cellular Camera is the ultimate remote scouting tool. The Reveal X Cellular Camera is not only small and discreet but powerful enough to handle the extreme outdoor weather. Tactacam’s REVEAL X Cellular Camera is the ultimate remote scouting tool. Start seeing the action as it’s happening even when you’re not in the woods! Data plans start as low as $4 per month, and the REVEAL boasts fast, easy setup and registration for added convenience.Tactacam’s Reveal X Cellular Camera is the ultimate remote scouting tool. It captures crisp 16MP daytime photos, and you can use the app to request HD videos and photos from the cam as needed. With a new built-in LCD screen and built-in GPS, its easier than ever to set up and track your camera. This makes the Tactacam Reveal X Pro Cellular Trail Camera the ultimate scouting tool. The REVEAL app offers advanced compression technology for higher image resolution, and allows you to view a summary your cameras in various locations to view all of your images on one screen. TACTACAM 2 Pack Reveal X Gen 2. See what's happening in the woods, even when you aren't there, with the Tactacam Reveal X Gen 2 Trail Camera Compact and durable, this game camera stands up to the elements while remaining discreet. The Tactacam Reveal X Pro Cellular Trail Camera is packed with even more features and benefits than the previous model. ![]() ![]() Hybrid Mode: Mix between the daily and instant settings check. Keep an eye on your land with the state-of-the-art Tactacam Reveal X Pro Cellular Trail Camera. Operating on 12 AA batteries (not included), this trail camera is LTE-enabled for wireless transmission of images and videos, straight to your smartphone or computer through the REVEAL app. TACTACAM Reveal X PRO Cellular Trail Camera, Verizon and AT&T, NO Glow, Integrated GPS Tracking, Built in LCD Screen, HD Photo and HD Video X-PRO + Tree Mount. The Tactacam Reveal X Pro is ideal for remotely and discreetly scouting your hunting property. Nighttime images are crisp and clear grayscale, while the day time photos are rich in color and clarity for optimal viewing. Featuring a lightning-fast trigger speed and an impressive 96' infrared flash, this trail camera captures crisp, HD images-day or night-without spooking wildlife. Never miss a shot with the TACTACAM® REVEAL™ Cellular Trail Camera. ![]() Not everybody at the marina is as equally thrilled about their teeth rattling as you may be. less loud thanks to the "captain's call" exhaust system allowing you to change from through the transom thunder to underwater rumble with the flip of a switch. Propelled by a supercharged fresh water cooled Mercruiser 496 with well over 600hp and tough Bravo One X drive with external hydraulic steering and high performance trim tabs this boat is all about speed, as measured by the Tattle-Tale GPS speedometer, and good looks, period.īut fast and loud is not the only choice, you've also got fast and. If you have a need for speed, this is the vessel indeed. In the world of racing pedigree boats you've got Fountain, Cigarette, Eliminator, Baja and more, among them stretching back to the early 60's is Checkmate, founded for racing and have always been about racing. See all 104 photos at www popyachts com/view/244849ġ987 High Performance Checkmate Convincor with 496 whipple super charger. Only 80 photos available on this website. If you're in the Philadelphia, Reading, Upper Darby, or Trenton areas, please use the form on this page to let me know what you're looking for and I'll get back with you today about any other high performance boat options we have locally that may meet your needs. We have several more Sea Ray, Scarab, and Fountain vessels for sale. This boat will be a great boat for the bay. This boat can get you, and three others where you want to be, enjoy the tunes on your pioneer stereo and enjoy the comfort of the pads that will keep you snug when you are motoring to your next destination. This boat has been well taken care of being cleaned and stored in covered storage after every use. The 1995 Checkmate Convincer cruises in the 50s MPH and can get up to over 70mph. On the dyno, Extension box with a bravo One drive!Īre you looking for a boat that will get you noticed with style and speed? Reason for selling is "Moving to Florida".Ģ8' Convincer with a 548 big block, 725 HP. See all 136 photos at www popyachts com/view/255111 ![]() Call us at (941) 914-9217 to discuss this vessel today. If you're in the Columbus, Newark, Mansfield, or Violet areas, please use the form on this page to let me know what you're looking for and I'll get back with you today about any other high performance boat options we have locally that may meet your needs. We have several more Sea Ray, Baja, and Formula vessels for sale. While we will always attempt to provide you with a TRUE representation of every vessel we market, during any purchase from Pop you will be encouraged to schedule an inspection from an independent, accredited surveyor and every purchase WILL ALWAYS BE subject to your satisfaction with the results from your independent survey and your own personal trial run. Used mainly for storage.ĭon't miss out on your opportunity to own this good condition Checkmate high performance! The cuddy could sleep 2, but would be a little tight. Newer upholstery along with the upgraded captain and passenger seats and stored indoors, this Checkmate boat can get you to your favorite spot at incredible speeds of up to 60 mph. She's been well-maintained, clean and ready to go. Powered by a 340hp OMC 5.7L King Cobra Big Block engine (rebuilt in 2020) will reach speeds of around 60mph. Located in Pataskala, Ohio, this 22ft 1989 Checkmate MAX229 high performance boat (freshwater cooling), freshwater use only and always stored indoors. ANY and ALL offers will be put in front of our seller! Submit your offer today!Įxcellent Condition Checkmate MAX229 With Ford 460 Cobra Big Block Freshwater Cooling 10hrs, New Ulphostrey and Seats!! In recent years, it has been aptly used to validate and study the size and relative abundance of small noncoding RNAs. However, it has the unique advantage of providing information about the expression level and size of the transcript so that it can be used to detect RNA degradation and alternative splice product of the same gene or repetitive sequence motifs, to reveal deletions or errors in transcript processing, and to isolate novel transcripts from heterogeneous mRNA pools. Therefore, Northern blotting is not a desirable method for RNA analysis. Furthermore, Northern blotting is a relatively labor-intensive technique because the procedure consists of several steps. Ĭompared to qPCR, the major limitations of Northern blot analysis are low detection sensitivity and easy RNA degradation by contaminated exogenous ribonucleases (RNases) in the course of extensive handling of RNA prior to blotting. However, its prominent advantage of higher detection sensitivity also condemns the risk of the publication of inconsistent, irrelevant, and even misleading data based mostly on flawed qPCR results and associated interpretation because of the difference in the quality of RNA samples and the efficiency of reverse transcription (RT), and the inappropriate methodologies selected for the normalization and quantification of data. Among these three methods, qPCR is an often used technology in current molecular biology laboratories due to its extreme detection sensitivity, high specificity and throughput. Northern blot analysis, ribonuclease protection assay, and real-time polymerase chain reaction (qPCR) are the three most commonly used techniques for studying the expression of one or a small number of genes. The multitiered technological methods used for identification of gene expression patterns can be divided into three categories: techniques for detecting the expression of one or a small number of genes, whole transcriptome-based approaches, and targeted next generation sequencing assays for the selected specific sets of genes or genomic regions. The study of gene expression can provide us with knowledge about gene function and regulation. The modification made and tips used ensured the efficient proceeding of the experiment and the resulting good performance, but without using special reagents or equipment. Using our modified Northern protocol, eight rounds of rehybridization could be performed on a single blot. We also report tips used in the modified Northern protocol, including the moderate concentration of formaldehyde in the gel, the accessory capillary setup, and the staining jar placed into an enamel square tray with a lid used for hybridization. Quantitatively controlled posthybridization washes combined with an ethidium bromide-prestaining RNA procedure to directly visualize prestained RNA bands at any time during electrophoresis or immediately after electrophoresis, which made the progress of the Northern procedure to be monitored and evaluated step by step, thereby making the experiment reliable and controllable. Successful detection of the low-expression gene using heterologous DNA probes in 20 µg of total RNA after a two-day exposure suggested an improvement in detection sensitivity. To maximize the retention of specifically bound radiolabeled probes on the blot, posthybridization washes were performed under only with moderate-stringency until the level of radioactivity retained on the filter decreased to 20~50 counts per second, rather than normally under high and low stringency sequentially for scheduled time or under only high stringent condition. ![]() In this paper, we report modifications and tips used to improve the traditional Northern protocol for the detection of mRNAs in total RNA. The Creative Commons Public Domain Dedication waiver ( ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. ![]() If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |