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Northern blot

The northern blot, or RNA blot, is a technique used in molecular biology research to study gene expression by detection of RNA in a sample. With northern blotting it is possible to observe cellular control over structure and function by determining the particular gene expression rates during differentiation and morphogenesis, as well as in abnormal or diseased conditions. Northern blotting involves the use of electrophoresis to separate RNA samples by size, detection with a hybridization probe complementary to part of or the entire target sequence; the term'northern blot' refers to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane. However, the entire process is referred to as northern blotting; the northern blot technique was developed in 1977 by James Alwine, David Kemp, George Stark at Stanford University, with contributions from Gerhard Heinrich. Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern.

The major difference is. A general blotting procedure starts with extraction of total RNA from a homogenized tissue sample or from cells. Eukaryotic mRNA can be isolated through the use of oligo cellulose chromatography to isolate only those RNAs with a poly tail. RNA samples are separated by gel electrophoresis. Since the gels are fragile and the probes are unable to enter the matrix, the RNA samples, now separated by size, are transferred to a nylon membrane through a capillary or vacuum blotting system. A nylon membrane with a positive charge is the most effective for use in northern blotting since the negatively charged nucleic acids have a high affinity for them; the transfer buffer used for the blotting contains formamide because it lowers the annealing temperature of the probe-RNA interaction, thus eliminating the need for high temperatures, which could cause RNA degradation. Once the RNA has been transferred to the membrane, it is immobilized through covalent linkage to the membrane by UV light or heat.

After a probe has been labeled, it is hybridized to the RNA on the membrane. Experimental conditions that can affect the efficiency and specificity of hybridization include ionic strength, duplex length, mismatched base pairs, base composition; the membrane is washed to ensure that the probe has bound and to prevent background signals from arising. The hybrid signals are detected by X-ray film and can be quantified by densitometry. To create controls for comparison in a northern blot, samples not displaying the gene product of interest can be used after determination by microarrays or RT-PCR; the RNA samples are most separated on agarose gels containing formaldehyde as a denaturing agent for the RNA to limit secondary structure. The gels can be stained with ethidium bromide and viewed under UV light to observe the quality and quantity of RNA before blotting. Polyacrylamide gel electrophoresis with urea can be used in RNA separation but it is most used for fragmented RNA or microRNAs. An RNA ladder is run alongside the samples on an electrophoresis gel to observe the size of fragments obtained but in total RNA samples the ribosomal subunits can act as size markers.

Since the large ribosomal subunit is 28S and the small ribosomal subunit is 18S two prominent bands appear on the gel, the larger at close to twice the intensity of the smaller. Probes for northern blotting are composed of nucleic acids with a complementary sequence to all or part of the RNA of interest, they can be DNA, RNA, or oligonucleotides with a minimum of 25 complementary bases to the target sequence. RNA probes that are transcribed in vitro are able to withstand more rigorous washing steps preventing some of the background noise. CDNA is created with labelled primers for the RNA sequence of interest to act as the probe in the northern blot; the probes must be labelled either with radioactive isotopes or with chemiluminescence in which alkaline phosphatase or horseradish peroxidase break down chemiluminescent substrates producing a detectable emission of light. The chemiluminescent labelling can occur in two ways: either the probe is attached to the enzyme, or the probe is labelled with a ligand for which the ligand is attached to the enzyme.

X-ray film can detect both the radioactive and chemiluminescent signals and many researchers prefer the chemiluminescent signals because they are faster, more sensitive, reduce the health hazards that go along with radioactive labels. The same membrane can be probed up to five times without a significant loss of the target RNA. Northern blotting allows one to observe a particular gene's expression pattern between tissues, developmental stages, environmental stress levels, pathogen infection, over the course of treatment; the technique has been used to show overexpression of oncogenes and downregulation of tumor-suppressor genes in cancerous cells when compared to'normal' tissue, as well as the gene expression in the rejection of transplanted organs. If an upregulated gene is observed by an abundance of mRNA on the northern blot the sample can be sequenced to determine if the gene is known to researchers or if it is a novel finding; the expression patterns obtained under given conditions can provide insight into the function of that gene.

Since the RNA is first separated by size, if only one probe type is used variance in the level of each band on the membrane can provide insight into the size of the product, suggesting alternative splice products of the same gene or re

Hossein Amanat

Hossein Amanat is an Iranian-Canadian architect. He is best known for being the architect of the Azadi Tower in Tehran, the Baháʼí Arc buildings in Haifa and the House of Worship in Samoa, he has been designated as the architect of the future Shrine of ʻAbdu'l-Bahá. As a young graduate from the University of Tehran he won a nationwide competition in 1966 to design the Shahyadd Tower, renamed the Azadi Tower in 1979; this first architectural project led to the opportunity to create some of Iran's most distinctive projects with reference to traditional Persian architecture. Amongst them are the initial buildings of the Sharif University of Technology in Tehran, Iran; the Persian Heritage Center, the Faculty for Business Management of the Tehran University and the Embassy of Iran in Beijing, China. As a member of the persecuted Baháʼí Faith, Amanat fled the country during the 1979 Iranian Revolution, he is International Studies at Yale University. Since moving to Canada in 1980, Hossein Amanat designed the three administrative buildings on the Baháʼí Arc in Haifa, the Baháʼí House of Worship in Samoa, the Jiang'an Library for the Sichuan University, the media library for the Beijing Broadcasting Institute.

He designed religious and cultural centers for the Baháʼí Faith near Dallas, Texas and Washington, D. C. several multifamily condominiums in Santa Monica and mixed-use high-rise buildings in San Diego and Burnaby, British Columbia, Canada. On 7 May 2019 the Universal House of Justice announced Amanat as the architect of the future Shrine of ʻAbdu'l-Bahá. 2001 American Concrete Institute Award 1995 Excellence in Building Design, Marble from Greece Competition 1985 Tucker Award for Architectural Excellence 1975 Royal Pahlavi Medal for Design 1971 Medal of Art - Iranian Ministry of Education List of historical Iranian architects Amanat Architect BBC interview with Hossein Amanat, October 2007 A list of Canadian Baháʼís in the news BBC interview: The man behind Tehran's Freedom Monument July 2009

Ridgeview Classical Schools

Ridgeview Classical Schools, or Ridgeview, is a free public K-12 charter school located at 1800 South Lemay, Fort Collins, Larimer County, United States. Ridgeview is chartered through the Poudre School District and serves 750 students, as of 2019; the school is accountable to its charter, the local school district, the state of Colorado. Grades K-8 follow a curriculum, inspired by the Core Knowledge Sequence. Since Ridgeview's inception in 2001, it has enriched this curriculum substantially. Elementary students continue it through eighth grade. Additionally, they begin studying Greek in third grade. Ridgeview begins teaching students cursive penmanship in third grade in a program, developed by Michael Sull and Marie Hornback. All elementary students study art and martial arts. Students in the elementary use the Riggs phonics system to learn to read and become proficient in diagramming sentences to improve their understanding of English grammar. Ridgeview's math program is influenced by the Singapore Math program, though elements of Saxon Math have been incorporated.

Students are ability-grouped at all grades in order to work at the highest level at which they can be successful across the curriculum. Character education is integrated throughout the curriculum. Grades 9-12 continue with a liberal arts curriculum. Ridgeview's high school courses make strong use of primary sources, where possible, Socratic discussion in classes. Students working towards a diploma take an equal number of courses in the humanities and natural sciences; this means four years of history, science and mathematics. Additionally, students are required to take courses in government and moral philosophy. Students are required to have a proficiency in Latin in order to graduate, they complete a 7,000-word senior thesis and complete a three-hour, written leaving exam. Students are required to defend it. Most students elect to take courses in music and art. Additionally, a variety of electives are offered, including not only AP courses, but concurrent enrollment courses through the University of Colorado.

A broad assortment of other electives are available to students ranging from neuroscience, computer programming, Russian literature, psychology of religion, along with many others. Modern language electives include: Spanish and German. All modern languages are taught by native speakers. U. S. News & World Report ranked United States high schools and awarded Ridgeview Classical Schools a gold medal, it is ranked 26th out of 458 high schools in Colorado, is ranked 28th among the nation's charter schools and 103rd among high schools nationally. ACT composite scores as reported by ACT from 2008-2013 are 26.3, 26.6, 26.2, 25.2, 27.2 and 25.8, with a state average during this time of 20.6. Ridgeview's Science Bowl team won the Rocky Mountain Regional competition in 2009, 2011, 2012, 2013. Ridgeview's K-9 chess team won Colorado State Championship in 2010 and 2011. In 2012 Ridgeview's K-9 chess team took fifth place nationwide; the high school chess team took third place in the state in 2012, first place in 2013, 2014, 2015.

In 2013, the K-3 and K-6 chess teams both took second place in Colorado. Ridgeview won CSU Math Day small school division in 2012. After School Club Chess Club Domino Club Engineering Club Folk Singers & Dancers Club Geography Club Hoplite Helpers Guitar Club Drama Club Science Bowl Scout Troops Madrigals Juggling Club Mock Trial Yearbook Club Veritas Student Council Student Ambassadors

Wallace House

Wallace House or Wallace Farm may refer to: J. N. Wallace House, Idaho, listed on the National Register of Historic Places in Ada County, Idaho Wallace House, Illinois Henry C. Wallace House, Iowa, listed on the NRHP in Madison County, Iowa Henry Wallace House, Des Moines, Iowa, NRHP-listed Charles Wallace House, Kentucky, listed on the NRHP in Ohio County, Kentucky Michael Wallace House, Kentucky, listed on the NRHP in Garrard County, Kentucky Samuel Wallace House, Kentucky, listed on the NRHP in Woodford County, Kentucky Wallace-Alford Farmstead, Kentucky, listed on the NRHP in Woodford County, Kentucky Napoleon Wallace House, Kentucky, listed on the NRHP in Green County, Kentucky Wallace House, listed on the NRHP in Boone County, Kentucky Everett Wallace House, Maine, listed on the NRHP in Washington County, Maine Wallace House, a National Historic Site Wallace House, listed on the NRHP in Laclede County, Missouri Wallace Farm, NRHP-listed Wallace House, NRHP-listed Jonathan Wallace House, New York, NRHP-listed Timothy Wallace House, New York, NRHP-listed Hambley-Wallace House, North Carolina, listed on the NRHP in Rowan County, North Carolina Charlton Wallace House, Ohio, NRHP-listed Wallace Farm, listed on the NRHP in Summit County, Ohio Wallace House, a fur trading station located in the French Prairie in what is now Keizer, Oregon Wallace-McGee House, South Carolina, NRHP-listed Gregg-Wallace Farm Tenant House, near Mars Bluff, South Carolina, NRHP-listed Wallace-Hall House, Texas, listed on the NRHP in Tarrant County, Texas Thomas Wallace House, Virginia, NRHP-listed Wallace-Jagdfeld Octagon House, Fond du Lac, Wisconsin, NRHP-listed Wallace House was an educator and politician in Newfoundland, Canada Gen. Lew Wallace Study, Indiana John M. Wallace Fourplex, Oregon Wallace-Baily Tavern, Brier Hill, Pennsylvania Wallace-Cross Mill, Pennsylvania Wallace Building Wallace

Darius Gaiden

Darius Gaiden is a shoot'em up arcade game and released by Taito in 1994. It is the third arcade installment of the Darius series. Darius Gaiden is a two-dimensional shoot'em up; the player controls a space ship named the Silver Hawk and must guide it through scrolling stages, destroying enemies and avoiding obstacles along the way. The ship is armed with forward-firing missiles, aerial bombs and a protective force-field, all of which can be upgraded by various power-ups that are dropped by specially-colored enemies when they are destroyed by the player. New to the Silver Hawk's arsenal in Darius Gaiden is the'black hole bomb.' When fired, the black hole bomb will create a large vortex in the center of the screen, which sucks in enemies and projectiles on the screen for a short moment while giving the Silver Hawk limited invincibility against enemies, until it explodes into a powerful ball of lightning that inflicts massive damage onto every enemy on the screen. Another feature introduced in Darius Gaiden is the ability to capture minibosses, who appear in every stage.

Each miniboss has a small, circular ball placed on them that, after receiving enough damage, will detach and float away, causing the miniboss to turn idle. If the player collects the ball, the miniboss will aid the player. After a brief period of time, or the player loses a life, the miniboss will explode. Shortly after the events of the first Darius, protagonists Proco and Tiat help refugees from the planet Darius flee from the destruction caused by the Belser Army, they find and set up a temporary colony on a planet named Vadis. Belser rises again and launches a surprise attack on a spaceport on Darius, being used to help ferry the remaining Darians to Vadis; the remaining refugees are killed, Belser sets course for Vadis. A fleet of Silver Hawks are sent to fend off Belser, but are annihilated. Once again and Tiat are given no choice but to fight off Belser once more. Darius Gaiden was ported to the Sega Saturn in 1995, to the PlayStation in 1996, to Microsoft Windows in 1997-2004; the Saturn and PC versions were released in Europe and North America by Acclaim and Interplay, respectively.

In 2006 it saw re-release on the PlayStation 2 as part of Taito Legends 2. The Saturn version of Darius Gaiden received positive reviews. Though most of Electronic Gaming Monthly's four reviewers felt the game was too hard, they praised the music, the absence of slowdown, most the striking graphics, recommended the game to shooter fans as "one of the most intense out there." GamePro's Air Hendrix complimented the controls, variety of enemies, menacing bosses and soundtrack, while criticizing the lack of selectable weapons, slowdown during busy moments, sound effects. He made particular note of the branching level layout, saying that it extends the longevity of the game to beyond that of a rental. Sega Saturn Magazine praised the multiple difficulty modes, high challenge and multiple paths through the game, their sole complaint was with the music, describing it as "some fat bint warbling away like an Old Spice advert." Despite being written nearly a year after the game's release and giving it a low score, Next Generation's brief review was relentlessly positive, describing Darius Gaiden as "A welcome blast from the past."

In a 2014 retrospective, Eurogamer called the game "one of the most confident and accomplished sprite-based games imagined". Studio " Ammonite " Official website Darius Gaiden at the Killer List of Videogames Darius Gaiden at MobyGames

American Airlines Theatre

The American Airlines Theatre the Selwyn Theatre, is a historic Italian Renaissance style Broadway theatre in New York City built in 1918. It was designed by George Keister and built by the Selwyn brothers. Used for musicals and other dramatic performances it was converted for film, it was used as a visitor's center but stood vacant for years until a 1997 renovation and restoration. It is located at 227 West 42nd Street. Named the Selwyn Theatre, it was designed by the architect George Keister and constructed by the Selwyn brothers and Archie, in 1918, it was one of three theatres they built and controlled on 42nd Street, along with the Apollo and the Times Square Theater. It was decorated in the style of the Italian Renaissance, had 1,180 seats. At the time of its opening, the design had several innovations, its most novel feature was separate smoking rooms for men and women. Additionally, each dressing room was equipped with a shower and telephone; the venue hosted major musical and dramatic productions, including Cole Porter's Wake Up and Dream in 1929, Three's a Crowd starring Clifton Webb in 1930–31, but became a cinema.

In the 1930s, 1940s and 1950s the theatre featured double features of standard Hollywood films. Beginning in the 1960s, until the theatre closed, the theatre featured grindhouse programs, it would return to legitimate theater several times over the next six decades, but fell into disrepair. It was used in the early 1990s as a home for the Times Square Visitors Center and for a limited production of Eugene O'Neill's The Hairy Ape, but for the most part, stood vacant; the City and State of New York took possession of the Selwyn in 1990. In 1992, it was one of six 42nd Street theatres to fall under the protection of the New 42nd Street organization; the Roundabout Theatre Company committed to renovating the Selwyn in 1997. It was restored to its former grandeur, renamed the American Airlines in honor of its principal sponsor, reopened on June 30, 2000; the American Airlines Theatre serves as the home of the Roundabout and houses its major dramatic productions. Productions that had more than 100 consecutive performances at the Selwyn Theatre: The Crowded Hour Tumble In Buddies Ed Wynn's Carnival Tickle Me The Circle The Blue Kitten Helen of Troy, New York Battling Butter |Battling Butter André Charlot's Revue of 1924 Kid Boots Charlot Revue Castles in the Air The Constant Nymph The Royal Family This Year of Grace Wake Up and Dream Three's a Crowd The Pajama Game achieved the box office record for the American Airlines Theatre.

The production grossed $477,030 for the week ending April 30, 2006. Broadway Theatres: History and Architecture, William Morrison, 1999, Dover Publications, ISBN 0-486-40244-4 Lost Broadway Theatres, Nicholas Van Hoogstraten, Princeton Architectural Press, 1997, ISBN 1-56898-116-3 Official website Spotlight on Broadway video history of the theatre; the Selwyn at the New 42nd Street Google Maps photo of entrance American Airlines Theatre at the Internet Broadway Database