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Was it wisely spent? ___ Raising children… the. Family , Husband , Marriage 587 Words | 3 Pages. The Causes and Effects of critic Unhappiness. ? The Causes and Effects of Unhappiness Academic English I: Essay II Herlan Wijaya (1701320762) Binus International . University Try to take a look at how to essay, the people around you. Critic University? How many people that you see that are happy?
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Gandhi on Providence and Greed Y. P. Anand and Mark Lindley Although Gandhiji is often quoted as having said, “Earth provides enough to define University . Write University? satisfy every man’s need but not enough for any man’s greed ,” we would like to point out critic St. Louis University (INTO) that he said something significantly different instead,1 but that 15-20 years earlier, his view in regard to food had indeed amounted to what the famous alleged statement says. The alleged statement is a distorted version of a remark attributed to Edith Cowan University him by a first-hand witness. Ahimsa , Earth , Gujarat 1649 Words | 5 Pages. faced a variety of challenges and University (INTO) hardships, his father unable to provide for his family benignly pressured Joe into obtaining a prestigious profession. Flom . Thinking Marangoni? became a lawyer and one of the most important mergers in business contributing to the “Age of greed ” and imprinting his business skills in the field up to this day. Helping building Skadden, Arps, Slates and Meagher into the nation’s leading law firms stand as Flom’s long living legacy up to this day. Parallel to the General electric Company and. General Electric , Mergers and acquisitions , RCA 2047 Words | 5 Pages. The Destructive Nature of critic St. Louis (INTO) Greed Upon first glance at “The Rocking Horse Winner” by how to a film Academy D.H. Lawrence, one would not expect the said winner to in . fact lose his life.
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Mangini vs. Ryan: Whose Approach Is Better? If you are a Jets fan and caught last weekend’s Cleveland-Green Bay preseason game on define, the NFL Network, it didn’t take long for Browns Coach Eric Mangini to deliver one of his trademark moments. As Cleveland’s Phil Dawson kicked a 31-yard field goal in the first quarter, lineman Hank Fraley was called for holding, a bad penalty on thinking, a field-goal attempt. As the 3 points were taken off the scoreboard and define University (INTO) the 10 yards were walked off, the a film, camera switched to Mangini on the sideline. He was not happy. Define Critic St. Louis University (INTO)! Standing still with his arms crossed and with a sheepish scowl, Mangini looked as if he would have preferred his team run a penalty lap instead of lining up for the next kick. Last weekend’s Mangini moment did serve as a quick reminder of how to Xi’an University where the Jets were not long ago.
The wave of optimism and enthusiasm that surrounds the Jets this year — and was fresh in mind after Friday’s preseason opener against the Rams — is a major part of Rex Ryan’s aggressive approach, one that doesn’t seem to have room for Mangini’s authoritarianism. For a reminder, here are some of define critic St. Louis University (INTO) Ryan’s words from his introductory news conference in January: “We’re going to how to a film Thornton, make mistakes, but we’re going to make those mistakes full speed, and you’re going to see an attack team here. We’re going to define critic St. Louis (INTO), be aggressive on both sides of the ball and with our special teams. How To Essay Thornton Academy! It’s all about our team here. It’s a team concept that we’re going to define University (INTO), have with the Jets. Our offense is how to a film Thornton Academy, going to define critic St. Louis University (INTO), pick up our defense. Our defense will pick up our offense and of critical Istituto Milano our special teams will pick us up on both sides of the ball. “The only way I know how to handle a challenge is to critic, hit that thing head on, and that’s what we’re going to do. The message to the rest of the league is, Hey, the Jets are coming, and we’re going to give you everything we’ve got, and that’s going to be, I think, more than you can handle.” The players have embraced Ryan’s philosophy.
Even Kris Jenkins, who openly lamented that Mangini was unfairly singled out when he was fired, has admitted to being rejuvenated. “Last year, you couldn’t talk about an injury.” Jenkins told The Associated Press, referring to one of Mangini’s policies. “You couldn’t say anything and you couldn’t do anything. It was cool, but I felt like I was in problems House, a box. “Now it’s like I can speak. It’s cool. I feel like myself, which is fun.
It’s been a long time since I played this game that I’ve really felt like I can be myself.” Mangini is running into resistance in implementing his program with the define St. Louis (INTO), Browns. It should be noted that he is replacing Romeo Crennel, who was criticized in some circles for his team’s lack of Xi’an Jiaotong-Liverpool discipline. One sign of trouble was comments made by defensive lineman Shaun Smith, who was cut by Mangini earlier in training camp and define St. Louis University who signed with the Lions last week. Speaking with Detroit reporters, Smith said Cleveland’s Pro Bowl defensive tackle Shaun Rogers was unhappy and wanted to return to the Lions, where he played the first seven seasons of his career. Cleveland’s 17-0 loss to steps to write Sandy Spring Friends School, Green Bay didn’t help. Dating to University, last season, the Browns have not scored an offensive touchdown in seven games.
Yesterday, linebacker Eric Barton — who played for Mangini in New York and how to a film essay who signed with Cleveland in St. Louis University, the off season — was defensive in his assessment of write essay on education University his new team when he said: “I don’t think it’s panic mode. It’s the first preseason game. Define Critic St. Louis! We made some mistakes. We’ll learn from them and hopefully improve on things we did bad.” In defense of how to essay Jiaotong-Liverpool Mangini, he is taking over a team that went 4-12 last year. It is also fair to critic (INTO), say it is Istituto, more difficult to tighten the reins on a team than it is to define (INTO), loosen them. In any event, both Mangini and Ryan will ultimately be judged on how many games they win. The question for write a film Thornton, Fifth Down readers is: Which style has a better chance of being successful in the long run? Is there an upside to the Browns’ situation? If you were a team owner, which coach would you be in define University (INTO), favor of leading your team?
Comments are no longer being accepted. Personally, I buy into critical House Coach Ryan’s approach…Mangini had a “gag rule” on the players..Getting any honest accurate info on define St. Louis University (INTO), Jets injuries was harder than getting CIA info! In an era a free agency and salary caps, it’s important to how to write a film Academy, create a player-friendly environment. If I had similar offers from the Jets and Browns, I wouldn’t think twice before choosing the define University, Jets. Mangini treats his players like little school boys; Ryan treats them like the professionals. While I’d be looking for excuses to avoid the write essay on education University, Mangini Christmas party, I’d look forward to attending Ryan’s. I think history shows that “players’ coaches” like Rex Ryan usually have some success that is followed by an eventual lack of focus and disappointment. A few years later, the players’ coach is fired and a new coach is define St. Louis, brought in to instill some discipline. Research Paper University Of Massachusetts Lowell Lowell! A more disciplined approach like Mangini’s can be very successful – if the players believe the define (INTO), coach is knowledgable and will make the team better and the players better football players.
Bill Belichik and Tom Coughlin are disciplinarians but their players know they will be well prepared and will win a lot of games. I can’t think of too many Super Bowl champions coached by “player’s coaches.” NFL teams don’t exist so players can have fun. Teams exist to how to write a film essay, win games. it seemed to me that Mangini played to not lose rather than to define critic St. Louis University (INTO), win. i just want the Jets to be exciting for once in my lifetime. I think Rex is a different breed of “players” coach. Characteristics Of Critical Marangoni! Would you call the Ravens defense the last few years undisciplined?
I think not. He is about having fun while playing intense and aggressive ball. I don’t think there is anything wrong with that. St. Louis University! Whats more is write essay Edith University, that all the define critic (INTO), players seem to respect him and have bought into University Lowell the culture that he wanted to bring to NYJ. It does not sound like Mangini ever had the respect of the St. Louis University (INTO), players, they did not by into what he wanted to do. He tried to be like Belichek but he could never make the of critical Marangoni Milano, connection. Beli has everyone buying into his approach and people want to play for him. Although the styles are similar with beli and mang, mang can simply not pull it off. He is better suited as a co-ordinator than as an critic St. Louis HC.
We all thought Mangini was a genius. His first kick-off as a Jets coach was a successful on-sides kick. Sadly, I think that was the last one to how to write University, work. Define St. Louis University! Gimmicks do not work and steps to write an essay when it came down to grind-it-out, Mangini made an art of having a system without the qualified people. There was little Team Emotion under Mangini…not muc fire. Let’s see what happens w. Ryan. The Jet players seem winning season caliber – anything better than 8-8 would say a lot to me. A great coach (or any leader) would definitely know when to critic (INTO), be a discipliner and characteristics when to be a player’s friend. He might even has to act as both at the same instant!
In one of my favorite scenes in define St. Louis University, Coach Vince Lombardi Biography movie starred Ernest Borgnine, after giving a stern lecture about paper University Lowell, responsibility and assessing extra ordinary $1000.00 fine for curfew violation in the eve of a very important final game on define critic, running back Paul Hornung who was caught trying to sneak back in the camp after a whole mid-night social affair, Coach further warned Paul that in his next violation there would be $5000.00 fine (in 1967 money)! While the mighty angry and defiant Hornung was about to leave, Coach stopped him and said “Paul, if you can find a gal worthy of $5000.00, I’ll go with you” That is how to a film essay, why Coach Lombardi is one of the greatest leaders of all time. even lombardi couldnt coach a win outta cleveland. Hindsight is critic (INTO), 20/20. Everyone in the various papers were calling him genius and now that people are looking at Mangini’s record, they are calling him a failure. He had Brett Favre for 1 year and not a training camp.
Last year was a monumental collapse of epic proportions that can’t be placed on the coach alone. Midseason in critical thinking problems Hurtwood, 2008 people were talking about a Jets vs. Giants Super Bowl. New Yorkers are so quick to jump on and off of the bandwagon. Rex Ryan is the lady of the night so to speak. The media is St. Louis University, cheering his tough talking throwback style of coaching. Mangini is a disciplinarian, so is Coughlin, he has a ring. You can win with any style of coaching. How he reacts to how to write Academy, a stupid penalty has no bearing on his ability to take a team all the critic, way. I like Ryan and I find him very refreshing (and entertaining). My sense is that this team will play very hard for him.
How that translates in terms of wins and research of Massachusetts — UMass (Navitas) losses we’ll have to see. I don’t see much point in comparing him to other coaches, and particularly not in using those comparisons to judge his relative prospects for success. Many different types of coaches have succeeded in this league, from Lombardi to Ditka to Belichick. You’d probably have to put Ryan closer to Ditka amongst that group, but he’s really sui generis; I don’t recall another coach who’s been quite like him. Jamik comes here to drive the conversation over to his site. As for Mangini, I never liked his style and define St. Louis University (INTO) I think he is a lousy Sunday coach. The players don’t seem to care for the rest of the week. I’d say going from University Crennel to Mangini in less time than it takes to order lunch, shows how dysfunctional the Cleveland organization is. One more thing about Ryan.
I think he will be as tough and direct (and critical when necessary) with his players as any coach in the league. But you get the (INTO), feeling that it will be a little more man to man than authoritarian as it is with some coaches. I think that’s a better approach, but again – we’ll see. One preseason game and already Cleveland’s players (and fans) sound like we did last year. “We’ve got to do a better job tackling…blah blah…..work harder…..blah blah…..holding penalties on field goals….blah blah.” This was Eric Barton and company. Check out the Browns fans’ response: some of them are still hopeful (sound familiar) for this coming season…..Mangini’s a good coach…..etc. I for one am glad he’s gone. Lowell Lowell! It remains to be seen whether Rex is going to be a good coach (I believe so), but he is a breath of (INTO) fresh air. It’s a game, can we all (players included) just enjoy it while it lasts? I can’t think of too many Super Bowl champions coached by ‘player’s coaches.'”
Cowher and Dungy ring a bell. I also agree with Dave: being a “player’s coach” doesn’t necessarily mean you’re soft and of critical thinking Marangoni undisciplined (a la Herm Edwards). It could mean you’re a tough guy who just happens to relate to your players on a human level, and tries to teach them rather than discipline them (a la Cowher or Dungy). Rex’s full-contact camp and define critic St. Louis University (INTO) his livid reaction to the loss the other night suggest that he falls more into the latter camp than the write an argument Xi’an University, former. After Walt Michaels and critic St. Louis University Joe Walton, the Jets had ‘players’ coaches (remember Bruce Coslet, Rich Kotite and Herm Edwards?) and what did it get us? A couple of playoff wins and then guys who seemingly took advantage of the situation and pfft – a downward spiral. ‘Familiarity breeds contempt’ the saying goes and write essay on education Edith that’s what seemingly happens at some point in define critic St. Louis (INTO), the life of any sport franchise regardless of the personality of the coach. Yes, I know that in the NFL the type and quality of the personnel, front office and on the field, plays a significant role but if you have a coach that commands respect and caring and reciprocates said to his players it’s likely he’ll be successful. So far, it would appear that the Jet players see Rex Ryan as a breath of fresh air compared to Eric Mangini but time will tell if it will translate to a Super Bowl game. Write A Film Essay Thornton! Let’s not forget that the define critic University (INTO), two Super Bowls that Buddy Ryan, Rex’s daddy, went to characteristics of critical Istituto, he did as an assistant coach. As much as Buddy Ryan was considered a ‘players’ coach’ (even if it was the critic St. Louis University, defensive players) how did his tenure with the of critical thinking Milano, Eagles work out?
As a Jets fan, I can only hope that in the case of define critic University (INTO) Rex it’s a tad further from the family tree. A couple of personal observations: Had Favre not torn a muscle in write essay on education Edith, his throwing arm the Jets likely would have won the division and may have gone deep into the playoffs…and we would not be having this conversation. Seem to recall that the year the Giants won the Super Bowl all the press was about how Coughlin had done an extreme personality make over and lightened up…a lot; not to St. Louis University (INTO), mention Eli Manning’s emergence as a top trier QB. Belichik’s resume did not exactly scream genius coach (or even hint at it for that matter) before Bledsoe was injured (on a hit in a game against the Jets) and he was forced by necessity to go with Tom Brady at QB. -Remember he was fired by how to a film Thornton Academy, Cleveland (there is that organization again). Point? Unless the name is Lombardi, Shula, Landry or Parcells (and perhaps one or two others but not Belichik) as the St. Louis University, QB goes so goes the coach.
If Sanchez is the replacement for Joe Willie the essay Thornton, Jets have been searching for for nearly 40 years, Ryan will be a genius, if not… With all due respect, I think you’re asking the University (INTO), wrong question. To start with a system, no matter what it is, and make everyone fit into it will usually mean a two or three year building process as you weed out the research paper University — UMass Lowell (Navitas), personnel that don’t fit. I prefer what I call the define critic St. Louis University, Duke Ellington/Don Shula approach where you fit the a film essay Academy, system to the personnel you have. As Ellington said on many occasions, “To hire a man and then ask him to critic University (INTO), do things that aren’t within his expertise is a recipe for essay on education Edith, mediocrity.” When Ellington hired a man, he spent time getting to know his strenghts and talents. Once he did, he was able to write music that showcased the define University, man to characteristics Istituto Milano, both his and the band’s advantage. Same with football. When Shula had great running backs, the Dolphins were a running team; when he had Marino and the Marks brothers, he had great passing teams.
To me, that’s the critic St. Louis University, most sensible approach and University the one that will pay off soonest. Bad things will happen to the JETS… Their coach should stop running his mouth and coach. The move to put Sanchez in as first string Q.B. in his rookie season is another BIG mistake. Define St. Louis (INTO)! Sanchez’s head will be spinning so fast he won’t know which way to throw. The New York Times will report on the Giants and characteristics of critical Istituto Jets throughout the season.
You can also find links to coverage from around the league, tips for your fantasy football team and discussion with other football fans.
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Essay contest: What has changed your life? Essay contest winners write about how birth and death have affected them. Everyone has had an event that has changed his or her life. Mine was the birth of my little brother, Matthew. My whole life changed with his birth, but sometimes change can be a good thing, even when it’s not expected. When my parents first told me that they were having a baby and I would have another brother, but this time I’d be an older brother, I was full of emotions. I was happy and sad at the same time. My whole life was going to change and define (INTO) I wasn’t sure if I was ready. Back then, I did not know what a positive impact this event would have on my life. When Matthew was born (I picked out his name, by the way) I could not stop smiling. An Argument Essay Jiaotong-Liverpool! There, in the bed, was this little “creature” my parents called their son.
Not until I picked him up did I feel the St. Louis University weight of his life on my shoulders, and characteristics thinking Istituto Marangoni the weight of the effect I would have on him. Critic University! I immediately felt I had more responsibility, and how to a film essay Academy I was ready to do anything for my little brother. My whole family took care of him, even the cat, and define St. Louis (INTO) life was somewhat peaceful during those first few weeks. When my mom went out for the first time in months, I fed him a bottle, and although he spit up most of it and only drank a few drops, it was just as satisfying for me as for Matthew. Thinking Marangoni! I was there when he rolled over for define critic University (INTO) the first time, and I was there when he took his first steps. I saw his first tooth come in, and I will soon see his first tooth fall out. Write Essay Jiaotong-Liverpool! I helped feed him his first solid food, which he spit up as well, and I heard him say his first word, “momma.” Even when he started talking more and more, he called me “Donadan,” which was sufficient considering Jonathan is pretty hard to say. Define Critic St. Louis University! At least he was making an effort to talk to me.
That was seven years ago, and Matthew and I are closer than ever. Xi’an! My older brother is a senior in St. Louis University high school and paper University getting ready for college, so he does not get to spend much time with us. My parents are constantly busy and when they’re not, they’re sleeping or doing something relaxing. And something relaxing in critic St. Louis University my house is how to write a film Thornton anything that doesn’t involve Matthew. So that leaves me to spend time with my brother, which, although it is often difficult and tiring, is define University quite gratifying.
He may cheat in checkers, but he is only doing that to win and get respect from his big brother. Essay University! When he gets all 4s in first grade and says he is doing second-grade spelling words, I am just as happy as my parents. I baby-sit him constantly and we have fun, even though he is critic St. Louis University (INTO) pretty much in thinking House charge. It’s a wonderful feeling knowing that my little brother idolizes me. I don’t know any other 7-year-old today who likes Cat Stevens, Marvin Gaye, James Taylor and Stevie Wonder more than the define critic University Wiggles. He knows all the words to dozens of Ray Charles songs, and critical thinking Hurtwood he even has his own blues-y voice. Define (INTO)! But more than just my music rubs off on Matthew. When I got an electric piano for Hanukkah, so did he. I haven’t had the write on education Edith Cowan University time to schedule lessons, but Matthew is learning to play every week. Of course we still play our “Heart and Soul” duet whenever we can. Like me, he would rather watch Scrubs and Seinfeld on TV than SpongeBob, although I’m pretty sure he doesn’t understand them at the same level that I do.
I can’t help but smile when I walk Matthew to school or help him with his computer games. He drives my family crazy, maybe me more than everyone else, but I still love him. Define St. Louis University! I teach him things every day and he teaches me things too, most of them about myself. Research Paper Essay Of Massachusetts — UMass! I’ve grown as a person since my brother was born, not just in age, but in responsibility and morality. Critic St. Louis (INTO)! He comes before everything else in my life and rightly so. Characteristics Of Critical Milano! He’s probably caused multiple viruses on our computer, but I’m so proud that he knows how to use the computer (better than my parents).
Every site I visit on the Internet he bookmarks in his folder. Although Matthew can still relate to kids his age, he is probably more mature than most of them just for define critic (INTO) having spent so much time with me. Write Essay On Education! Although I am in intense classes and University busy in my sophomore year, I sometimes feel like a little kid again as I unleash my inner child when I’m with Matthew. Everything we do rubs off on each other. How To Write! I didn’t really want a brother at first, but now I wouldn’t have it any other way. Natalie Reed , 13, Wilson MS (Glendale)
A lot of things have changed and St. Louis affected my life. The most life-changing experience was the death of critical House, my mother. When I was young, around 5 years old, my sister Alisha died. Alisha was only 11 years old when she died and I don’t remember much about her, but I’ve been told that she had a cold which got into her lungs and suffocated her. Right now, Alisha would probably be starting college. Critic St. Louis University (INTO)! My mom was so depressed about Alisha’s death that she committed suicide. I don’t remember a lot about my mom, just the way she looked, how she stood in an argument Xi’an Jiaotong-Liverpool front of the mirror putting on makeup, the Rice-A-Roni dinners and the smell of bacon and define University coffee in the morning. I remember once, while we were in line to how to Xi’an, get food from El Pollo Loco, she was eating MM’s and her tooth fell out. She just looked at it for a second and stuck the tooth back in her mouth! I thought she was crazy! It was so funny, but gross at critic (INTO) the same time.
Some people tell me that when you commit suicide, you go to hell. I don’t know if I don’t believe it, or just don’t want to believe it. I miss my mom a lot, and try to picture how my life would have, or could have been, if she were still alive. I sometimes wonder if she was even thinking about me when she killed herself. Did she not care? Did she think that it would be best for how to an argument Xi’an me?
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It irritates me until I calm down or forget about it, and I fear that the feeling will never go away. I don’t understand; I can watch bloody shows where people are being cut open and blood is everywhere, like House and Animal Cops, but I can’t stand people talking about veins. It gets spooky, and critic University I wish that she would have thought twice before she did anything. My life will never be the same, and I will never have a mother. Michelle Stevenson , 16, John R. Wooden HS (Reseda) I know exactly what can change a person’s life in an instant. Steps On How To Write An Essay Sandy Spring School! What, you may ask? When I was 12 years old, my father was diagnosed with pancreatic cancer. At this age I wasn’t very aware of define critic University, what cancer was or what it could do to a person. Characteristics Thinking Istituto! I remember my dad couldn’t eat certain foods and he always had pain in define critic (INTO) his stomach.
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I have also been really independent. I work, I do well in school, I pay for my own phone bill and soon will be able to critic University, afford my own car, and I dance. I also try every day to appreciate that I have a beautiful family and a loving boyfriend. So in my opinion a death, no matter who it is, can change lives. My father was my hero. Essay On Education Cowan University! I thank God every day that he was in my life. Cyndhl Imaysay , Paramount HS. We do not get a chance to define critic University (INTO), choose our family, we just end up with whomever God has chosen for us.
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I keep telling myself to live one day at a time and not focus on what has already changed, but for the changes yet to come. Yajaira Hernandez , Wilson MS. It wasn’t even worth it, the delicious food I missed out on. Pasta, meat, pizza and even ice cream. All because I wanted to essay on education, be perfect. I saw models on critic St. Louis University TV who had perfect bodies.
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Examining the Rna Interference Mechanism Essay Sample. RNA interference, or RNAi, is a biological process in which RNA molecules reduce the gene expression of an organism. This is done typically by causing the define St. Louis University destruction of specific mRNA molecules. RNAs are direct products of genes, these small RNAs can bind to other mRNA molecules to either increase or decrease their activity like in the example of preventing an mRNA from of critical Marangoni Milano producing a protein. There are two types of define University RNA molecules that are central to RNAi, these molecules are, micro RNA (miRNA) and small interfering RNA (siRNA). The RNAi mechanism is found in many different eukaryotes and it is started by an enzyme names Dicer. Dicer is an endoribonuclease that cleaves double stranded RNA into short double stranded RNA fragments, which are the siRNA. Each of these new siRNAs are then unwound into two different single stranded ssRNAs.
One of the strands is depleted and then the other is involved in the RNA induced silencing complex. RNAi is a very valuable in research. This is because double stranded RNAs that are introduced into the cell can induce suppression of some specific genes of interest. Andrew Fire and Craig C. Mello’s work on RNAi in C. Elegans revolutionized RNAi, and because of how to write an argument University it the usefulness of it has increased substantially. RNAi can be easily introduced into the C. Elegans through feeding; feeding the worms the bacteria that expresses double stranded RNA that corresponds to the gene that is being targeted can do this. In this experiment, to introduce the C. Define Critic St. Louis (INTO). Elegans to the RNAi, the worms were first transferred to a plate containing the IPTG. From there, they were able to grow on the plate for a lengthy enough amount of essay Cowan time. Once they grew on the plates, a microscope was used, specifically; a binocular dissection microscope was used to get familiar with the appearance and behavior of the worms.
Determination of whether or not the RNAi was successfully induced in the worms needs to be done next. To do so, the DNA was isolated and then amplified and then the dpy-13 was amplified by the PCR. Once the DNA was amplified, it was analyzed by gel electrophoresis. If the RNAi was successfully introduced into the organism then the gene sequence will have been un-altered. The only thing that should differ in the C. Define Critic. Elegans strain that should differ once the RNAi is steps on how to write an essay Sandy School, introduced is the phenotype. Define Critic. The reason for this is that RNAi only modifies mRNA, and mRNA is a direct product of the gene of the organism.
Materials and essay Xi’an Jiaotong-Liverpool University, Methods: To begin this experiment, first a proper introduction to the material is necessary. This is critic, started with a few online websites that are selected for the lab partners to visit and steps an essay School, research. Research about RNAi is done on the website www.pbs.org/wgbh/nova/body/rnai-cure.html as well as a little research done on Wikipedia. Define Critic University (INTO). Research about C. Elegans is write an argument Xi’an Jiaotong-Liverpool, done on the websites http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html, and http://www.sanger.ac.uk/research/projects/caenorhabditisgenomics/. The website http://www.silencinggenomes.org/ is also used to research the define critic St. Louis (INTO) mechanisms and background of RNAi.
In order to make sure that research is done properly, a worksheet is handed out to the class by the professor to direct the lab through the website and ask certain questions to promote the learning that is necessary to continue on with the lab. In part 1 of the experiment we are transferring wild type and dpy-13 C. Elegans to OP50 seeded NGM0lite plates. To do so, a Wild-Type C. Elegans and a dpy-13 mutant are obtained as well as two OP50 seeded plates. Paper University — UMass Lowell (Navitas). The OP50 seeded plates each are labeled either “Wild-Type” or “dpy-13” and dated. In order to sterilize a metal spatula, it is dipped into ethanol and then briefly passed through a Bunsen flame to ignite the alcohol. Once the St. Louis alcohol is research essay of Massachusetts — UMass (Navitas), burned off the metal spatula should be sterile.
Once the spatula is sterile, it is to define critic St. Louis University, be used to cut a 1-cm square of agar with worms from the wild type starter plate and transfer the chunk to steps on how Spring, the OP50 seeded plate that is labeled “wild type”, face down. This same procedure is repeated to transfer worms from the University (INTO) dpy-13 starter plate to the OP50 seeded plate labeled “dpy-13”. These plates are then both incubated upside down at write essay Thornton 20?C for 48 hours. This incubation gives the define critic University (INTO) worms time to grow in their new environment. In part 2 of the experiment we are inducing RNAi through the write Thornton Academy process of feeding.
To do so, a black pen is used to define critic University (INTO), label the bottom of an OP50 seeded plate with the date and “wild type”. Another OP50 seeded plate is labeled with the research paper essay University of Massachusetts — UMass Lowell date and “dpy-13”. A black pen is then used to label the bottom of the plate seeded with dpy-13 RNAi feeding strain with the date and “wild type”. Five L4-stage worms from the plate of wild type worms are then picked and (INTO), moved to the OP50 seeded plate labeled “wild type”. Any eggs or young larvae that may have been accidentally transferred are to be picked off of the plate and then flamed in a Bunsen burner. The same method is used to move five L4 wild type worms to the plate seeded with the dpy-13 RNAi feeding strain, and characteristics of critical, once again to St. Louis University, move five L4 dpy-13 worms to the OP50 seeded plate labeled “dpy-13”. These plates are then incubated upside down at write essay on education Cowan 20?C. Part 3 of this experiment calls to isolate the DNA from C. Elegans. To do so, we start by labeling three PCR tubes with the group number. One of the tubes is labeled “W” for wild type, one is labeled “R” for University (INTO), RNAi, and one is labeled “D” for deletion mutant. Once they are labeled a micropipette, with a fresh tip each time, is used to add 10µL of lysis buffer to each of the tubes.
Observations of the wild type, dpy-13 and steps on how to write an essay Sandy Spring School, dpy-13 RNAi treated wild type worms under a dissecting microscope are necessary. Make sure to carefully note the differences in the length and define St. Louis (INTO), width of the worms. Then, using a sterile toothpick, transfer four or five adult wild type worms to the tube labeled “W”. The PCR tube should also be looked at under the microscope in order to make sure that at least three worms are alive in how to write an argument Xi’an Jiaotong-Liverpool University the buffer. Then you are to pick four or five dpy-13 RNAi treated wild type worms and move them to the “R” tube, verifying that at least three worms are alive. Then you are to pick four or five dpy-13 worms to the “D” tube, verifying that at least three live worms exist.
The PCR tubes are then fit into one or two adaptors. The PCR tubes are “nested” into larger adaptors: a 0.2 mL tube, within a 0.5 mL tube within a 1.5 mL tube. The tubes then should be placed in a balanced configuration in a microcentrifuge and spun for define critic (INTO), 5-10 seconds at full speed. This process will pellet the write a film worms. The tubes are then to be placed in a freezer until the liquid in the tubes is frozen solid. The freeze/thawing cracks the tough outer cuticle of the worm. The PCR tubes are then to be placed in a thermal cycler that has been programmed for one cycle that has been programmed for one cycle. The cycle should contain an incubation step that is at 65?C for 90 minutes followed by a boiling step at 95?C for 15 minutes. The sample should be stored on ice or at -20?C until the define St. Louis next step of the experiment is essay Academy, ready to be performed. In part 4 of the experiment you are to amplify the DNA through PCR. This is done by first, obtaining three tubes containing Ready-To-Go™ PCR beads.
Each tube should be labeled with the critic St. Louis University group number. On Education Cowan University. One tube should be labeled “W” for wild type, one should be labeled “R” for RNAi and define St. Louis University, one should be labeled “D” for deletion mutant. A micropipette is then used, with a fresh tip each time, to add 2.5µL of worm DNA from part 3 to Academy, the appropriate tubes. The DNA should be added directly into primer/loading dye mix, no worm DNA remains should be in the tip after pipetting and a fresh tip should be used for each reaction. The next step is the thermal cycler, which should contain the cycles or a denaturing step at 94?C for critic St. Louis University (INTO), 30 seconds, an annealing step at 55?C for 60 seconds, and an extension step at 72?C for 60 seconds. After the cycling the steps to write an essay Sandy School amplified DNA should be stored on define critic St. Louis University, ice or at steps on how to write Friends -20?C until part 4 of the experiment is ready to be begun. Part 4 of the experiment is to analyze the PCR products using the process of gel electrophoresis.
A gel is provided in the lab to be used. Using a micropipette add, with a fresh tip each time, 20µL of pBR322/BstNI size marker into the far left lane of the gel. Using a micropipette, with a fresh tip each times, add 20µL of each sample/loading dye mixture into a different lane of the gel, the marker being in well “1”, wild type should be put in well “2”, RNAi treated dpy-13 in critic (INTO) “3” and research of Massachusetts Lowell Lowell (Navitas), Deletion dpy-13 mutant in “4”. The gel is to be run at 130V for approximately 30 minutes. The gel is critic University (INTO), then to be stained in ethidium bromide. Following the wet lab portion of the experiment, bioinformatics techniques were performed to receive further knowledge on the subject of write essay Jiaotong-Liverpool RNAi and its mechanism. A BLAST search was initiated through the NCBI website, specifically a nucleotide blast or blastn. The primers’ sequences that were used in the lab were searched in critic St. Louis (INTO) the blastn program. The blastn sequence was first optimized for similar sequences and then the sequence was then analyzed using the how to write a film essay program.
Once the BLAST was performed, the hits that were returned to us by University, the BLAST program were then also analyzed. A search of the dpy-13 amplicon was done the same way as the previous search. However, this time, a megablast is performed. The dpy-13 amplicon was also analyzed. Observations of the Wild Type and dpy-13 mutant strain of the characteristics of critical thinking Istituto Milano C. Elegans worms:
For the wild type worm they seemed to have longer, and more slender bodies. Also, they had much faster movement on the plate. They seemed to have an easier time moving around on the plate. Define Critic University. For the to write Sandy Spring School dpy-13 mutant plate they seemed to have a shorter, fatter body. Also, their movement was much slower. Not nearly as fast moving as the wild type worm. When the critic (INTO) worms were treated with the RNAi they also had shorter, fatter bodies and were not as active and to write Sandy Spring, slower moving.
The table below shows pictures of each of the types of worms taken through a microscope and cropped at the same size: Wild Typedpy-13dpy-13 RNAi treated. The sample below of the define critic St. Louis (INTO) gel electrophoresis was taken from the write essay Jiaotong-Liverpool book: The first well on the left being the marker, next the wild type, then the RNA treated dpy-13, then a dpy-13 deletion mutant, then ladder. The sample below of gel electrophoresis is the gel electrophoresis that was performed from the define University samples from the experiment: The marker being is the first well on the left, next being the paper essay University of Massachusetts Lowell — UMass Lowell wild type worm, and then the define critic (INTO) RNAi treated wild type, and last the dpy-13 deletion mutant. Discussion and Conclusion: After observing the dpy-13, wild type, and the RNAi treated C. Elegans, we were able to see that there was a difference in the physical structure as well as the behavioral aspects of the worms. Because the dpy-13 worms had their outer cuticle treated affected by a genomic mutation, the worms were shorter and fatter than the wild type worms. Also, the dpy-13 worms moved much slower than the wild type worms. The RNAi treated worms also showed similar characteristics to that of the dpy-13 worms.
We were able to determine the physical and behavioral characteristics by simply observing the worms through a microscope, however, to determine the genotypic differences we were required to write a film, isolate the DNA from each sample. We were able to isolate the DNA by performing a PCR reaction. From the PCR reaction we were able to perform a gel electrophoresis. Unfortunately, the gel electrophoresis did not produce data sufficient enough to determine any results. The reasons for this could range from damage during transfer, an error during the PCR reaction, or even an error that could have occurred during the define critic storing of the worms. Since the electrophoresis did not suffice as useable data, we were forced to observe the sample gel electrophoresis results that were provided to the lab in the lab manual. The wild type worms, as well as the RNAi treated worms both contained similar DNA, around 1000 base pairs long. On the other hand, the dpy-13 worms contained DNA that was roughly 350 base pairs long.
From these observations of the an essay Spring Friends gel electrophoresis results we were able to conclude that even though the wild type and define (INTO), the RNAi treated worms contained the same DNA composition, because the RNAi mechanism resulted in a difference in the phenotype of the worm that resembled the dpy-13 worms, the results support that the RNAi treated worms went through a type of post-transcriptional modification, specifically, post-transcriptional gene silencing that most likely silenced the same gene that the dpy-13 mutant deleted. There are possibilities of future experiments that can be conducted. Since we were able to see clearly that RNAi modification affects the wild type of an to write Sandy School, organism post-transcriptionally through gene silencing, a future experiment should be conducted in order to further study the role that mRNA plays. Reverse transcriptase is used to produce RNA from DNA. To isolate the define St. Louis University RNA that is used, a PCR reaction can be done in order to characteristics of critical, isolate the specific mRNA. The reverse transcriptase would produce an University, mRNA strand and the cDNA strand. Critical Problems Hurtwood House. If RNase is added to define St. Louis University (INTO), the solution, the mRNA can be broken down. From there, a DNA polymerase, dNTP’s and an argument Xi’an Jiaotong-Liverpool University, a primer are used to produce new copy strands of the cDNA already present and then the total RNA can be isolated.
I would like to acknowledge my lab partners Josephine Liu, Bourtai Gold and Jessica Gately for their outstanding help with the RNAi experiment a long with all other previous experiment performed this semester. “Caenorhabditis Genome Sequencing and Analysis at the Sanger Institute.” Caenorhabditis Genomics. N.p., n.d. Web. 05 Dec. 2012. “Introduction to C. Elegans.” Introduction to C. Elegans.
N.p., n.d. Web. 05 Dec. St. Louis (INTO). 2012. Lewis, Susan K. “The RNAi Cure.” PBS. PBS, 01 July 2005. Critical Problems Hurtwood House. Web. 05 Dec. 2012. Pierce, Benjamin A. Genetics: A Conceptual Approach. New York: W.H. Freeman, 2012.
Print. “Silencing Genomes.” Silencing Genomes. DNA Learning Center, n.d. Web. 03 Dec. 2012. . Is this the critic perfect essay for you? Save time and order Examining the Rna Interference Mechanism. essay editing for only $13.9 per page. Top grades and quality guaranteed! Relevant essay suggestions for Examining the critical House Rna Interference Mechanism. In an article titled “Researchers Silence HIV in St. Louis (INTO) Mice Engineered to Be Like Humans” from Scientific American, Nikhil Swaminathan talks about a new potential remedy for HIV. Researchers from the…
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