Term Paper: Transcription Is a Process

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SAMPLE EXCERPT:

[. . .] The termination complex consists of terminal RNA and mRNA that associate with polypeptide chains that had hydrolyzed

Question 4 There are 64 codons total. Why do you suppose some amino acids have only one or two codons while others have more? What is the wobble?

Genetic redundancy is a condition in which an individual suffers from a condition where a certain biochemical function cannot work as it is impaired by one or two genes. This condition makes the amino acids have only one or more codons. There are 4 bases and 21 amino acids. Four to the power of one gives one, 4 to the power of 2 gives 16, 4 to the power of three gives 64. The situation cannot work with the two bases because it would create a shortage of amino acids and this is unhealthy for the body. Thus, the body uses three bases to cater for the 21 amino acids.

In case of a point mutation inside the genome, the process may not actually mature to the protein getting mature. A perfect example is in the case ACC that has the responsibility of coding Threonine. If a person was to get ACA, the result would remain the same and the reason for this is that it would still code Threonine. This is where explanation of the wobble hypothesis comes in.

The Wobble Hypothesis states that the pairing of basis relaxes at the third position. The reason for this is so that a base can have the ability to pair up with another base other than the original ones. In some RNA, anticodons bear inosine at position three. Inosine has the ability to pair with three specific codons and thus a human being or any other organism does not require having over 60 RNA molecules, even 30 are enough (Singer, 2011)

Question 5. Detail the coliform test. What are the acceptable values and what do levels indicate?

There are different ways in which to detail the coliform test. Fecal coliforms comprise part of a larger group of coliforms that come from the origin of felicons in general. Human feces however have a number of aerogenes and other substances that are separate from the original coliform group and are termed as non-fecal segments. To differentiate between coliform and other products of the fecal origin, temperatures must be set at 39 degrees and higher. This is necessary for incubation to take place effectively. Fecal coliform can ferment carbohydrates at a temperature of 44 degrees in a time span of 24 hrs. It is important to leave the mixture unattended for that number of hours. The product that one attains after that is the total chloroform that does not ferment in that temperature (Campbell, 2009).

In order to enumerate fecal chloroform, one can go about this in two ways. The MPN (Most Probable Number) is one of the ways and it involves serial dilution in which there is observation of one bacterium instilled in a fermentation tube with conditions that support its survival. For this analysis to go perfectly, one requirement is five tubes of dimensions that do not exceed three decimals. Before acting on the tubes, the fermentation tube should portray gas production or lack of production. This is necessary in the determination of the initial number of bacteria that was in the tube initially.

The main aim of having the dilution is for the analysis to observe tubes that would produce gas eventually and the one that would not produce any gas. It is important to calculate the water to use when carrying out the dilution process. This might seem as a rather unnecessary step as it determines the width of the tubes to use during the distillation. Wastewater treatment water and can effectively work with dilutions of between 1 and 0.01ml. This is because the water is within 200/100ml of chlorinated effluents discharge and these are acceptable levels.

Question 6. Explain how to clone Eukaryotic genes. Include as many details as possible.

In the contemporary world of science, researchers carry out their work in order to advance knowledge and apply personal development. One of the most important aspects to acknowledge is the presence of Eukaryotic cells that are some of the latest microorganisms such as fungi and yeast. Modern scientists have even come up with ways through which they can clone these cells (Singer, 2011).

The most common method that cloning of these cells takes place is through structural gene sequences. Through this process, a complementary DNA and RNA receive transcribing through in vitro. After this process, they both become double-stranded. The procedure takes place inside the Escherichia coli plasmids by the tailing technique. Coli transformants that come from the DNA contain globin sequences which attainment is through hybridization of RNA to DNA caused by clones developed and then lysed in situ. This process takes place on nitrocellulose. In order to estimate the amount of globin sequence inserted coming from the DNA, fingerprint analysis of the RNA sequence combined to become a hybrid of purified chimeras takes place (Singer, 2011).

An alternative procedure of the same experiment states that when carrying out the cloning, the scientist does not recommend the use of DNA. This is because introns cannot receive splicing from prokaryotes. Instead of using DNA, c-DNA is the most appropriate one to use. The c-DNA comes from m-RNA through reverse transcription. As the m-RNA does not have its introns present, the c-DNA also lacks, this procedure however makes the process longer. Both procedures are correct but when choosing which the best to incorporate into an experiment, it is important to understand the type of product undergoing cloning.

Question 7. Explain how Polymerase Chain Reaction (PCR) works. Why is it such a powerful tool in diagnostics and other molecular biology applications?

Polymerase chain reaction is a technique in molecular genetics used in construction multiple genes in the process of gene sequencing. With the new technology, the DNA builds the gene copies and makes many copies from one copy that is the sample. The work of PCR is amplifying the gene to million copies and allowing for gene identification using the visual techniques. Gene identification depends on the size and type of charge of the DNA. The PCR is a powerful tool used in laboratory procedures such as in mapping techniques and clinical techniques for fingerprinting, detecting bacteria and viruses (Sun, 2012).

How does PCR work?

When amplifying a DNA segment using the PCR technique, the first step is heating the sample until the DNA denatures or separating the sample to two single strands of DNA. The 'Tap polymerase' enzyme forms two new DNA strands from the original strands, resulting in duplication of the original DNA. Each new molecule has an old and a new strand of DNA. Each of these strands form two new copies and the process continues leading to a cycle of denaturing and synthesizing of the new DNA. This repetition forms many similar copies of the initial DNA segment. The whole PCR cycling process is mechanical, and it completes within a few hours, directed by the thermocycler machine. This machine increases the temperature in the reaction in order to allow the synthesis and denaturing of the DNA. The PCR leads to the formation of million copies of a single DNA sequence by amplification of the strand.

Question 8. Describe how real-time PCR works.

The real-time PCR, well-known as the RT-PCR is a technique of RNA detection used in the TaqMan and SYBR Green applications. SYBR green offers the easiest and most economical method in the detection and quantitative of the PCR products during real-time reactions. The application binds the DNA in double strands and emits light upon excitation, increasing the fluorescence in the PCR product. The SYBR Green application has its advantages such that is sensitive, easy to operate and inexpensive. On disadvantages, it can bind any DNA with double strands such as primer-dimmers or other non-specific products of the reactions. They can cause in overestimation of the target concentration. The SYBR Green works well in the single PCR products containing primers with proper designs and the non-specific background exists in later cycles. This application is economical for detection of real-time products (Clark, 2013).

The TaqMan Probes application relies on fifth nuclear activity in the DNA polymerase. The PCR techniques use this application in increasing specificity of real-time PCR assays. TaqMan probes are oligonucleotides with dye, fluorescent in nature attached at the 5' end and a quencher moiety at the 3' end. The figure below shows the basic appearance of a TaqMan probe.

The probes hybridize the external regions of the PCR products. During the unhybridized nature the flu or proximity and that of the quench molecules prevents detection of fluorescent signals from the probe. After replication of template by the polymerase, the TaqMan probe bounds on the template, and the nuclease activity in the 5' section… [END OF PREVIEW]

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