Richard Remick Smothers is an American actor, comedian and musician. He is half of the musical comedy team the Smothers Brothers, with his older brother Tom. Smothers was born in the son of Ruth, a homemaker. Smothers, an Army officer who died as a prisoner of war in April 1945. After moving to Southern California, Dick attended Verdugo Hills High School in Tujunga and graduated from Redondo Union High School in Redondo Beach and attended San José State University called San José State College. At SJSC, Smothers participated as a distance runner for the track team; the Smothers Brothers have appeared on numerous television shows over the past three decades, including two shows of their own: The Smothers Brothers Show, a sit-com from 1965 to 1966. In 1977, he appeared twice. In 1993, he played one of the characters on cartoon Christmas movie Precious Moments: Timmy's Special Delivery. Without Tom, he appeared in the 1995 Martin Scorsese-directed film Casino in an uncharacteristically serious role as a dishonest Nevada State Senator.
His character and the dialogue in one scene was based on the career of former United States Senator Harry Reid, who once chaired the Nevada Gaming Commission. In February 2010, Smothers filed for Chapter 11 bankruptcy protection. In May, he and his brother announced their retirement from touring. Smothers has been active in both road racing and drag racing, he is the father of six children: Dick Jr. Andrew, Sarah and Remick, he resides in Sarasota, Florida. Dick Smothers on IMDb Smothered: The Censorship Struggles of the Smothers Brothers Comedy Hour on IMDb Dick Smothers at The Interviews: An Oral History of Television
The versatility of polymerase chain reaction has led to a large number of variants of PCR. Only a small modification needs to be made to the standard PCR protocol to achieve a desired goal: Multiplex-PCR uses several pairs of primers annealing to different target sequences; this permits the simultaneous analysis of multiple targets in a single sample. For example, in testing for genetic mutations, six or more amplifications might be combined. In the standard protocol for DNA Fingerprinting, the targets assayed are amplified in groups of 3 or 4. Multiplex Ligation-dependent Probe Amplification permits multiple targets to be amplified using only a single pair of primers, avoiding the resolution limitations of multiplex PCR. Multiplex PCR has been used for analysis of microsatellites and SNPs. Variable Number of Tandem Repeats PCR targets areas of the genome; the analysis of the genotypes of the sample involves sizing of the amplification products by gel electrophoresis. Analysis of smaller VNTR segments known as short tandem repeats is the basis for DNA fingerprinting databases such as CODIS.
Asymmetric PCR preferentially amplifies one strand of the target DNA. It is used in some sequencing methods and hybridization probing, to generate one DNA strand as product. Thermocycling is carried out as in PCR, but with a limiting amount or leaving out one of the primers; when the limiting primer becomes depleted, replication increases arithmetically through extension of the excess primer. A modification of this process, named Linear-After-The-Exponential-PCR, uses a limiting primer with a higher Melting temperature than the excess primer to maintain reaction efficiency as the limiting primer concentration decreases mid-reaction.. Some modifications are needed to perform long PCR; the original Klenow-based PCR process did not generate products. Taq polymerase can however amplify targets of up to several thousand bp long. Since modified protocols with Taq enzyme have allowed targets of over 50 kb to be amplified. Nested PCR is used to increase the specificity of DNA amplification. Two sets of primers are used in two successive reactions.
In the first PCR, one pair of primers is used to generate DNA products, which may contain products amplified from non-target areas. The products from the first PCR are used as template in a second PCR, using one or two different primers whose binding sites are located within the first set, thus increasing specificity. Nested PCR is more successful in amplifying long DNA products than conventional PCR, but it requires more detailed knowledge of the sequence of the target. Quantitative PCR is used to measure the specific amount of target DNA in a sample. By measuring amplification only within the phase of true exponential increase, the amount of measured product more reflects the initial amount of target. Special thermal cyclers are used. Quantitative Real-Time PCR methods use fluorescent dyes, such as Sybr Green, or fluorophore-containing DNA probes, such as TaqMan, to measure the amount of amplified product as the amplification progresses. Hot-start PCR is a technique performed manually by heating the reaction components to the DNA melting temperature before adding the polymerase.
In this way, non-specific amplification at lower temperatures is prevented. Alternatively, specialized reagents inhibit the polymerase's activity at ambient temperature, either by the binding of an antibody, or by the presence of covalently bound inhibitors that only dissociate after a high-temperature activation step.'Hot-start/cold-finish PCR' is achieved with new hybrid polymerases that are inactive at ambient temperature and are only activated at elevated temperatures. In touchdown PCR, the annealing temperature is decreased in cycles; the annealing temperature in the early cycles is 3–5 °C above the standard Tm of the primers used, while in the cycles it is a similar amount below the Tm. The initial higher annealing temperature leads to greater specificity for primer binding, while the lower temperatures permit more efficient amplification at the end of the reaction. Assembly PCR is the synthesis of long DNA structures by performing PCR on a pool of long oligonucleotides with short overlapping segments, to assemble two or more pieces of DNA into one piece.
It involves an initial PCR with primers that have an overlap and a second PCR using the products as the template that generates the final full-length product. This technique may substitute for ligation-based assembly. In colony PCR, bacterial colonies are screened directly by PCR, for example, the screen for correct DNA vector constructs. Colonies are sampled with a sterile pipette tip and a small quantity of cells transferred into a PCR mix. To release the DNA from the cells, the PCR is either started with an extended time at 95 °C, or with a shortened denaturation step at 100 °C and special chimeric DNA polymerase; the digital polymerase chain reaction amplifies thousands of samples, each in a separate droplet within an emulsion. Suicide PCR is used in paleogenetics or other studies where avoiding false positives and ensuring the specificity of the amplified fragment is the highest priority, it was described in a study to verify the presence of the microbe Yersinia pestis in dental samples obtained from 14th Century graves of people killed by plague during the medieval Black Death epidemic.
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Asheik Jarma was elected Governor of Borno State, Nigeria in October 1983, holding office until the military coup on 31 December 1983 that brought General Mohammadu Buhari to power. He was elected on the National Party of Nigeria platform. In the lead up to restoration of democracy in 1999, Jarma was a founding member of the People's Democratic Party. In April 2001 the PDP suspended Jarma from its board of trustees for one month for flirting with other political associations. In November 2001 he was a member of the interim contact and mobilisation committee for the newly formed United Nigeria Democratic Party. In July 2008 a Senate ad hoc committee probing a food crisis implicated Jarma among others for abandoning £11.4 million worth of silo contracts. In October 2009 he denied that he had attended the kick-off meeting of the launched National Democratic Movement and said he had always been a bona fide member of the PDP. In December 2009 Jarma endorsed the PDP candidature of Ambassador Saidu Pindar for Borno Governor in the 2011 elections