Reverse transcriptase

several complications, prompting virus detection methods to develop rapidly. (Chen, 2), (CDC) Materials and method: The detection of the ZIKV using reverse transcriptase-PCR begins with a probe and primers that were designed specifically for this experiment. FD3/FU1 forward and reverse primers were sequenced from the NS5 region that had a size of 1083bp. The primers were then amplified using plasmids and then extracted via QIAquick gel that separated them based on size. A QIAamp was then used to extract the Zika virus’s RNA. A 100 mL of a synthetic sample that contains the virus was obtained to perform the next portion of the experiment. The sample was mixed with the AVL-Carrier RNA buffer, and a 400 mL of ethanol was added after 10 minutes. The mixed sample was placed in a silica column and centrifuged for 1 minute. The mixture was then washed with AW1 and AW2 buffers. Finally the extracted RNA was eluted with TE buffer and kept at -800C. (Faye 1,2) The method began by using a positive and a negative control. The positive control was from the virus’s nucleic acid and the negative control was water. The reagents that were employed in this method and the amplifying process of the viral RNA used were included in the Quantitect One-Step RT-PCR kit. 25uL of extracted RNA sample was utilized in this technique while 0.25uL of reverse transcriptase RT and Taq polymerase were added with 10ul of 2 x Quantite RT-PCR buffer. Two 1.25 uL of primers were added as well with the…

Objective: We started by isolating a DNA plasmid from a bacterial culture for cloning. Then, we reverse transcribed mRNA to amplify it in a polymerase chain reaction. Finally, we amplified a specific gene (actin) through PCR. Protocol: A. Isolation of Plasmid DNA from a Bacterial Culture: We were provided with a tube containing 1 ml of bacterial culture. We centrifuged it for 5 minutes at 10000 g. Then, we removed the supernatant by pouring it off and we used a small chem wipe to remove any…

resistance; developing new drugs and diagnosing tests can help treat the evolving bacteria and identify just exactly how it is evolving (CDC, 2015). 2. We have come a long way in the treatment and control of the spread of HIV. In a few sentences answer the questions below. Site sources. The NIH site on HIV will have most of the information you will need to answer the questions. Answer each part in 2-3 sentences. (2 points) NIH: http://www.niaid.nih.gov/topics/hivaids/Pages/Default.aspx You can…

containing the target DNA sequence to be amplified, 2 primers (forward and reverse), heat-stable. PCR is similar to DNA sequencing. This is based on a reaction where a primer and dNTP’s are added with a DNA template and DNA polymerase (Taq) 1. The difference in PCR is that if you use a primer on the 5' end it will make a new direction and gene. The primer is made in the opposite strand and the direction which is called lagging strand. The template is melted, the primers are annealing then two…

BigDye® Terminator v3.1 Cycle Sequencing Kit provides everything we need in order to generate a sequence all we need to do is add one primer and the DNA template. These reagents are suitable for performing fluorescence-based cycle sequencing reactions on single-stranded or double-stranded DNA templates, on polymerase chain reaction (PCR) fragments, and on large templates. First, you must prepare your template by performing a DNA extraction and purifying your template. The purification will be…

In the play Helen by Euripides the main character, Helen often finds herself in complicated situations. To get out of those situations, Helen believes that her only option is to manipulate, deceive, and lie to people. Helen’s main goal is to be reunited with her husband, Menelaos, in her homeland. Helen persuades characters to do things her way by manipulation, lies, and deception. Throughout Helen, the main character manipulates Teucros, Menelaos, and Theoclymenos using deception, persuasion,…

The unconstitutional use of racial quotas in the admission process of the medical school in the University of California at Davis, opened one of the most famous court cases that had to be settle in the Supreme Court of the United States of America. The case involved the admission practices of the Medical School of the University of California at Davis which reserved 16 out of 100 seats in its entering class for minorities, including Blacks, Chicanos, Asians, and American Indians. The purpose of…

In 1968, the Medical School of the University of California at Davis established a racial quota, according to the Regents of the University of California v. Bakke (1999). The university’s admissions office was run under two separate programs: “the regular admissions program and the special admissions program.” Minority students were admitted through the special admissions program and had different requirements to fulfill than the students admitted through the regular admissions program (1999).…

The We Are Done Movement came about in response to similar protests at Universities around the country. It is a way for this group to express their opinions on social issues present on the campus of the University of Alabama, but many people don’t agree with the organizations ideas. Joe McDonald wrote an essay depicting why the We Are Done Movement is wrong and displayed evidence behind his reasoning. Joe argued that: the safe space this organization is calling for is only making the divide…

to assist in admission policies and increase diversity on college campuses by providing equal access to education for women and minorities. While affirmative action does sound good on paper, some believe that the affirmative action program is outdated and needs to be abolished because it causes reverse discrimination “by favoring one group over another,…