Restriction Enzyme Analysis of DNA

Restriction Enzyme Analysis of DNA Agarose gel electrophoresis is a method used in biochemistry and molecular biology. It is used to separate DNA or RNA molecules by size. Since DNA and RNA have negatively charge, when they go through an agarose matrix with an electric field, they will move from cathode to anode. The shorter molecules move faster and migrate farther than longer molecules; therefore the different sizes molecules can be separate. The most important factor which affects the migration is the length of the DNA molecules. A restriction enzyme, is an enzyme that cuts double stranded DNA following its specific recognition of short nucleotide sequences, is used to cut the DNA into small fragments. The unique recognition sequences are usually tetra- or hexanucleotide palindromes with axes of dyad symmetry. Which means the sequence on one strand reads the same in the reverse direction on the complementary strand, e.g. GTATAC and its complementary strand CATATG. Recognition sequences in DNA differ for each restriction enzyme, producing differences in the length, sequence and strand orientation of the DNA fragments. Plasmid is an extra chromosomal DNA molecule, which is capable of replicating independently of the chromosomal DNA. It is circular and double stranded. Plasmid is usually found in bacteria. The size of plasmid is between 1 to 200 kilobase pairs. In this experiment, a specific recombinant plasmid pBR325 was analyzed, and hence a restriction map would be constructed. METHOD As described in the practical manual RESULTS From the photograph, four unknown plasmid fragments migrated distance could be measured and hence the four fragments size could be found: In lane 2: migrated distance of the fragment was 36.5mm, therefore 11.00 kilobase would be the size of this fragment; In lane 3: migrated distance of the near fragment was 42.5mm, therefore 7.08 kilobase would be the size of this fragment; migrated distance of the further fragment was 66mm, therefore 3.98 kilobase would be the size of this fragment; In lane 4: migrated distance of the near fragment was 40.75mm, therefore 7.94 kilobase would be the size of this fragment; migrated distance of the further fragment was 73mm, therefore 2.82 kilobase would be the size of this fragment; In lane 5: migrated distance of the near fragment was 50mm, therefore 6.31 kilobase would be the size of this fragment; migrated distance of the further fragment was 53mm, therefore 5.62 kilobase would be the size of this fragment; The 4 unknown plasmid size and migrated distance were showed in following table: The size of the plasmid could be found, which was about 11 kilobase. Since the plasmid was only cut by EcoRI + BamHI in about 4:8 ratio of the plasmid, therefore the restriction map for EcoRI + BamHI could be construted: DISCUSSION The nucleic acids migrated from cathode to anode since it had negatively charge. The cutting ratio of three enzymes could be found by the size of fragment as result showed. The last lane, which was the uncut plasmid, was loaded, in order to compare with the EcoRI enzyme cut fragment. The migrated distance of this uncut plasmid was large than EcoRI cuts migrated distance, which could be measured from graph. EcoRI cuts fragment was like uncoiled elastic. It encounters more resistance migrating through a gel because it is spread out and will be in direct contact with more of the gel matrix. Uncut plasmid was tightly coiled, like a balled up elastic. While the molecule may be the same size, the coiling compresses it, allowing it to encounter less resistance when migrating through the gel. This indicated the minimum base pairs of the plasmid. If a hybrid recombinant plasmid was constructed from pBR325 by the insertion of a fragment of DNA at the BamHI restriction site, firstly the total size of the plasmid got bigger. To determine the size of the inserted fragment, EcoRI and PstI were used to cut the plasmid, and result was compared with original plasmids fragment to get the size of the inserted fragment. There was no additional band visible behind the main uncut plasmid band in lane 7. If there ware, they must be the chromosomal DNA bands, the absent of these in lane 2, 3, 4 and 5 was because they were too small, might not be seen, or ran off the gel. The fluorescent bands of the DNA restriction fragments in lane 2, 3, 4, and 5 were several fold brighter than the uncut plasmid band in lane 7. It was because that the DNA fragment are free at both ends and can uncoil and pick up as much EtBr as it fits, whereas the uncut plasmid could not uncoil more than a certain amount without the phosphate chain breaking: for every incorporated molecule of EtBr it had an area of local under-coiling that had to be compensated by another area of over-coiling, this area would not incorportate any EtBr. So the number of EtBr molecules it could pick up was limited because of sterical considerations. Therefore the DNA fragment had less sterical restrictions, stained more than the uncut plasmid.

Consumer Behaviour in the Unisex Shoe Store Category :: Marketing, argumentative, persuasive

This report analyzes the role personal, social, and situational factors in consumer behaviour in the unisex shoe store category. Observations were made at three different shoe retailers including Town Shoes, Payless Shoe Source, and The Shoe Company. These three stores offer very diverse retail environments targeted towards different demographics of people. This report also found that the retail setting, such as music, light, and organization, had an effect on the actions of the consumers in the store. The time of day also had an effect on consumer’s purchasing behaviour. Environmental analysis Town Shoes: The store had bright lights and loud pop culture music without lyrics which attracted a younger demographic of customers. There were â€Å"Buy one get one half off† signs displayed all around the store, which encourages consumers to buy more than one pair of shoes. The shoes are displayed without any form of organization; brand name shoes are mixed in with non-brand low quality ones, all stocked in piles on the floor. There are no specific try-on areas for shoppers to try on their shoes and the sales people were mostly unavailable to answer questions because of the high turnover rate of customers entering and exiting the store. The sale racks that post 50% have mixed shoes of different color, style, and size. Most of the shoes are for the current season (i.e. sandals, pumps) and the majorities are female sandals and heels. Payless: The sale discount signs and cheap price of shoes is the big attention grabber for this store. The medium lighting with soft background music provides consumers a relaxing environment to shop. The shoes are displayed in a warehouse style according to size and gender and accessories. There are also medium quality bags and accessories located near the cashier that satisfies consumer’s desire to take part in the summer fashion. The store layout implies that there is no other inventory since all the shoes are out on display. Most of the shoes only targets the current season. There are limited numbers of try-on areas at the end of the aisles near the back of the store, encouraging shoppers to test out shoes. The employees do not seem to approach customers to help them, but are available for questions. The Shoe Company: The recognizable designer shoes are on the display to attract consumers who seek for quality shoes or the desire to identify themselves as having a higher social status.

Final Exam Critical Thinking Essay

Our newest product Wet Wonders lip gloss, has not been performing very well. Creative Cosmetics needs to come up with different ways to market our lip gloss in order to boost sales. The projected sales for Wet Wonders lip gloss have slumped and we are in danger of having to scrap this product and its promotional campaign. During product development, our initial testing proved that Wet Wonders can be successful based off the positive feedback we received. In order to boost sales, Creative Cosmetics needs to analyze the role of advertising in society, why it’s so important and how to effectively use it. Without advertising consumers would not be told what to buy. Advertising is the reason our homes are filled with so many items, why we drive the cars we do and why we wear the clothes we wear. There have been many studies on the increase of advertisements over the years. â€Å"In 1915, a person could go entire weeks without observing an ad. The average adult today sees some three thousand every day† (Twitchell 203). Advertising also fuels the need to shop which in turn stimulates the economy. Economist can track consumer spending every quarter allowing them to gauge how well the economy is doing. Due to budget constraints, Creative Cosmetics chose to only sell Wet Wonders through ads on the radio. Using only radio ads to market our product, has not proven to be very effective. While radio allows you to tell a story and take the listener’s mind on a journey, it is not the most effective means of marketing a lip gloss. The radio paints a broad stroke, meaning it is heard by many different people. There are specific times when we could have our ads running that could pinpoint a certain group of people, although it’s not necessarily guaranteed. Another downside to radio ads is they don’t have the ability to allow the consumer to see what they are missing. It’s not enough to say we have a wonderful product. We want to make the consumer feel that their lives would be so much better with our lip gloss and if they don’t buy it then they are missing out on something great. In other words, we have to convince them that they need our lip gloss in order to make their lives as exciting as the woman wearing it. Television could do that most effectively. Television commercials would give us the visual component we need. Our ads could really drive home the message that Wet Wonders lip gloss can change your whole look in instant. We want women to feel as though they can go from drab to fab with just one swipe of our lip gloss. Television would give us the ability to make women as though they would feel, sexy, confident and desirable when they wear our lip gloss. We could run our commercials during peak times that women are watching television such as the morning and afternoons. While there are a lot of women working outside the home, there are still a good number of women that stay at home to raise families. An ideal time to run our commercials would be during talk and game shows as well as soap operas. Another idea would be to use a famous person or celebrity in our advertising. People like to feel as though they trust the person they are buying something from and even though they may not personally know the celebrity they trust them. Since we know our product would be best suited being marketed on a visual platform, we should also consider magazine ads. Magazines would be a perfect way to market our Wet Wonders lip gloss. With magazine ads we could saturate the market with our product and message. There are teen magazines, young adult beauty magazines, and magazines for the more mature woman. Every one of these genres have women that are sexy, confident, and desirable. This is exactly the woman we want to market to. Magazines have the ability to make consumers visualize themselves as that sexy, vibrant woman that wears Wet Wonders lip gloss just by looking at a picture.

Classroom Reflection - 1256 Words

My experience was informational. I was able to see how my students are inside the classroom environment with my teacher how they are in art. I was also able to observe another classroom and see how they differ from my class. All of these are completely different spaces where the children act differently. Comparing these different environments provided a unique insight into how these children act differently depending on who is around. Having been with these children a lot now, I have the ability to interact with them and get to know them individually. This helps in my classroom assessments because I am able to differentiate between how children normally behave and what the normal classroom routine is like. When the children were in my†¦show more content†¦One thing that I don’t like about my class, though, is that there are no rules posted. I know in the other class, there are rules posted so when a child goes against the rules, they can refer back to the list. 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