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,…

How Reverse Mortgages Work: Calculate Your Reverse Mortgage and Decide What Best Suits You There are two basic considerations before one can qualify for a reverse mortgage. First, he/she should be at least 62 years old. Secondly, this individual needs to own a home which he/she is expected to live in. Furthermore, he/she is required to receive client information from counseling services that are Federal-approved before receiving the mortgage loan. An online reverse mortgage calculator lets you…

recognize it. Policies that have been implemented to counter this discrimination have recently been decried as going so far as to implement “reverse discrimination” because of the benefits they inure to those who have historically been harmed at the expenses of the majority. Reverse discrimination occurs when those in power discriminate against members of the majority group in favor of the minority group (Lerner & Nagai, 2000).This type of discrimination includes policies such as preferential…

is of a hate sticker against Islam with razors on the back so that those who try to remove it could possibly get hurt while attempting to do so. Even with affirmative action, also known as “positive discrimination,” minorities face racism. Affirmative action, first established in 1961 (“A Brief History”), was to compensate for the racism that minorities faced in the past, mostly stemming from slavery. Since then, different propositions have been made to alter the meaning of affirmative action…

different opinions on the view of affirmative action. Edwin C. Hettinger is on the side calling affirmative action “reverse racism” itself suggests that it is discrimination: discrimination towards…