Plasmid Extraction :: essays research papers

Introduction   Ã‚  Ã‚  Ã‚  Ã‚  Chitobiase, from Vibrio harveyi, is a membrane bound lipoprotein involved in the degradation of chitin. Chitobiase is similar to and may share a common ancestry to the a-chain of human b-hexos-aminidase. Chitobiase is encoded by chb.   Ã‚  Ã‚  Ã‚  Ã‚  In this experiment, a restriction map for restriction enzymes Eco R1, Pst1 and Hind III using Southern hybridization and restriction analysis of pRSG 192. pRSG 192 is a recombinant plasmid derived from the chb gene and pUC 19, a 2.7kb engineered plasmid which encodes for ampicillin resistance, a portion of the lac operon and a multiple cloning region . The chb gene exists as a 3.6 kb insert in the mutiple cloning region of pUC 19.   Ã‚  Ã‚  Ã‚  Ã‚  The major goals of Experiment One will be to isolate pRSG 192 from an overnight culture of E. coli, amplify a region of the chb gene using PCR, and to map restriction sites within the chb gene using restriction analysis and Southern hybridization. Methods Plasmid Isolation Four microfuge tubes containing cell pellets representing 3.0ml of cells(2 x 1.5ml) from an overnight culture of E. coli were prepared. The supernatant fluid was discarded and each pellet was resuspended in 150ul of TE buffer(10mM Tris-HCl, pH 8.0; 0.1 EDTA). 300ul of SDS(1% SDS, 0.2 N NaOH) was added to each pellet. The tubes were placed on ice for five minutes, after which, 225ul of ice-cold 3M potassium acetate(pH 4.8) was added. The tubes were again placed on ice for five minutes and subsequently microfuged for five minutes. The supernatants were recovered and transferred to new tubes. One volume of phenol/chloroform was added to each new tube. The tubes were shaken vigorously for two minutes and centrifuged for five minutes. The upper, aqueous phase was recovered and transferred to a new tube. One volume of chloroform was added to each tube. The tubes were vigorously mixed and microfuged for three minutes.

Conflicts in the Godfather Essay

The Godfather is an insightful sociological study of violence, power, honor and obligation, corruption, justice and crime in America. Part I of The Godfather Trilogy centers on the Corleone crime â€Å"family† in the boroughs of New York City in the mid 1940s, dominated at first by the aging godfather/patriarch â€Å"Don† Vito Corleone. As a turn-of-the-century Silician immigrant, he is the head of one of the five Italian-American â€Å"families† that operates a crime syndicate. The ‘honorable’ crime â€Å"family,† working outside the system due to exclusion by social prejudice, serves as a metaphor for the way business (the pursuit of the American dream) is conducted in capitalistic, profit-making corporations and governmental circles. Although conflict in life may seem unpleasant, literature readers find it to be quite exciting. The characters involved within a conflict clearly stand out as to what they truly believe in and who they are as a person. In Mario Puzo’s, â€Å"The Godfather†, a number of conflicts are revealed through the Mafia underworld. The first, and most important conflict that is demonstrated in â€Å"The Godfather†, is revealed right away through â€Å"The Assassination Attempt on Don Corleone†. The Don shows the conflict of man vs. society by first refusing to enter the drug business. This decision, will ultimately effect the future conflicts that are revealed throughout the story. By refusing to participate in the drug business, the Corleone Family becomes outcasts of the five major crime families in New York. By doing so, the Don sparks the war between his family and the other five families. This war ends up lasting for several years and costs many lives. The second conflict that is present in â€Å"The Godfather†, is the conflict of divergent ideas vs. his father’s ideas. Santino, â€Å"Sonny†, Corleone, demostrates this conflict by deciding to go against his father’s beliefs. Sonny feels that drugs are the thing of the future and he decides that if his father dies, he will make the deal of entering the narcotics business. Tom Hagen, the adopted son of Vito Corleone, also believes that by entering the drug business, the Family will become much more successful later down  the road. A third conflict that is present in â€Å"The Godfather†, is the conflict of man vs. himself. This conflict is demonstrates through the character of Michael Corleone, the youngest son of the Don. Michael’s conscience is at war with his with his desire. After the assassination attempt on his father, Michael feels that he needs to get involved and help his father continue the Family business. However, his mind also feels that it is wrong to enter the criminal business. In conclusion, it is clear that many conflicts exist in Mario Puzo’s, â€Å"The Godfather†. They include, man vs. society, divergent ideas vs. father’s, and man vs. himself. Together, these conflicts create an intriguing plot that captures the viewer’s attention to the Mafia Underworld during the 1940’s and 1950’s in New York. Mario Puzo’s materpiece studies the power of conflicts that involve violence, power, honor, obligation, justice, and corruption.

Weeks v. United States The Case and Its Impact

Weeks v. U.S. was a landmark case that laid the basis for the exclusionary rule, which prevents illegally obtained evidence from being used in federal court. In its decision, the court unanimously upheld Fourth Amendment protections against unwarranted searches and seizures. Fast Facts: Weeks v. United States Case Argued: Dec 2—3, 1913Decision Issued:  February 24, 1914Petitioner:  Fremont WeeksRespondent:  United StatesKey Questions: Could the items obtained without a search warrant from Mr. Week’s private residence be used as evidence against him, or was the search and seizure without a warrant a violation of the Fourth Amendment?Unanimous Decision: Justices White, McKenna, Holmes, Day, Lurton, Hughes, Van Devanter, Lamar, and PitneyRuling: The Court held that the seizure of items from Weeks residence directly violated his constitutional rights, and also that the governments refusal to return his possessions violated the Fourth Amendment. Facts of the Case In 1911, Fremont Weeks was suspected of transporting lottery tickets via mail, an offense against the Criminal Code. Officers in Kansas City, Missouri, arrested Weeks at his work and searched his office. Later, officers also searched Weeks home, seizing evidence including papers, envelopes, and letters. Weeks was not present for the search and officers did not have a warrant. The evidence was turned over to the U.S. Marshalls. Based on that evidence, the Marshalls conducted a follow-up search and seized additional documents. Prior to the court date, Weeks’ attorney petitioned the court to return the evidence and to prevent the district attorney from using it in court. The court denied this petition and Weeks was convicted. Week’s attorney appealed the conviction on the basis that the court had violated his Fourth Amendment protection against illegal searches and seizures by conducting an unwarranted search and by using the product of that search in court. Constitutional Issues The main constitutional issues argued in Weeks v. U.S. were:Whether it is legal for a federal agent to conduct an unwarranted search and seizure of a person’s home, andIf this illegally obtained evidence can be used against someone in court. The Arguments Weeks’ attorney argued that officers had violated Weeks’ Fourth Amendment protections against unreasonable searches and seizures when they entered his home without a warrant to obtain evidence. They also argued that allowing illegally obtained evidence to be used in court defeats the purpose of the Fourth Amendment. On behalf of the government, attorneys argued that the arrest was based on sufficient probable cause. The evidence uncovered in the search served to confirm what the officers had suspected: Weeks was guilty and the evidence proved that. Therefore, the attorneys reasoned, it should eligible to be used in court. Majority Opinion In a decision delivered by Justice William Day on February 24, 1914, the court ruled that the search and seizure of evidence in Weeks home violated his Fourth Amendment right. Fourth Amendment protections apply to someone whether accused of crime or not,† according to the Court. Officers needed a warrant or consent to search Weeks home. The federal government also violated Weeks Fourth Amendment protections when the court refused to return evidence seized during an unreasonable search. In finding that the search was illegal, the court rejected one of the governments main arguments. The governments attorneys had attempted to show the similarities between Adams v. New York and Weeks case. In Adams v. New York, the court ruled that evidence incidentally seized while conducting a legal, warranted search may be used in court. Since officers had not used a warrant to search Weeks home, the court refused to apply the ruling reached in Adams v. New York. The Justices ruled that the illegally seized evidence was fruit from the poisonous tree. It could not be used in a federal court. Allowing the district attorney to use such evidence to convict Weeks would violate the intent of the Fourth Amendment. In the majority opinion, Justice Day wrote: The effect of the Fourth Amendment is to put the courts of the United States and Federal officials, in the exercise of their power and authority, under limitations and restraints as to the exercise of such power and authority, and to forever secure the people, their persons, houses, papers, and effects, against all unreasonable searches and seizures under the guise of law. The Court reasoned that allowing submission of illegally obtained evidence actually encouraged officers to violate the Fourth Amendment. In order to deter violations, the court applied the exclusionary rule. Under this rule, federal officers who conducted unreasonable, unwarranted searches could not use the evidence they found in court. The Impact Prior to Weeks v. U.S., federal officers were not punished for violating the Fourth Amendment in pursuit of evidence. Weeks v. U.S. gave the courts a means of preventing unwarranted intrusions on a person’s private property. If illegally obtained evidence could not be used in court, there was no reason for officers to conduct illegal searches. The exclusionary rule in Weeks only applied to federal officers, which meant that illegally obtained evidence couldn’t be used in federal courts. The case did nothing to protect Fourth Amendment rights in state courts. Between Weeks v. U.S. and Mapp v. Ohio, it was commonplace for state officers, unbound by the exclusionary rule, to conduct illegal searches and seizures and hand the evidence to federal officers. In 1960, Elkins v. U.S. closed that gap when the court ruled that the transfer of illegally obtained evidence violated the Fourth Amendment. Weeks v. U.S. also laid the groundwork for Mapp v. Ohio in 1961, which extended the exclusionary rule to apply to state courts. The rule is now considered a fundamental element of Fourth Amendment law, providing the subjects of unreasonable searches and seizures a unified manner of recourse. Weeks v. U.S. Key Takeaways In 1914 the court ruled unanimously that evidence obtained through an illegal search and seizure could not be used in federal courts.The ruling established the exclusionary rule, which prevents the court from using evidence that officers uncover during an illegal search and seizure.The exclusionary rule only applied to federal officers until Mapp v. Ohio in 1961. Sources Root, Damon. Why Courts Reject Illegally Obtained Evidence.  Reason, Apr. 2018, p. 14.  General OneFile.http://link.galegroup.com/apps/doc/A531978570/ITOF?umlin_m_brandeissidITOFxidd41004ce.Weeks v. United States, 232 U.S. 383 (1914).