Its name was coined from one of its key processes i.e. DNA polymerase. A DNA template is produced and the chain reaction that follows is the concluding part of the process.
A major requirement of recombinant DNA techniques is the "availability of large quantities of specific DNA segment" (Bastianutto et al 2006). Cloning which was the preferred method involves transformation of a plasmid vector into bacteria that is then cultured. The cloning process is not as efficient as PCR in terms of amplification of DNA. In addition PCR can allow the generation of millions of copies of DNA from a single or few pieces.
PCR occurs in stages, the Initialization, Denaturation, Annealing, Extension/elongation, Final elongation. The Initialization step involves heating to 94-96C. After the heating stage, Denaturation produces a single DNA strand. This is caused by breaking the hydrogen bonds between DNA strands, thus causing the melting of the DNA template and primers. Temperature is lowered during the annealing step. Here primers are bound to the single stranded DNA template by the polymerase. At the Extension/elongation the DNA polymerase produces a new DNA strand by adding dNTPs. The last PCR cycle is the Final elongation, the step is done to ensure that all single stranded DNA are fully extended. ...Show more
This essay will examine the Polymerase Chain Reaction (PCR) technique by taking a closer look at its emergence as the preferred technique for multiplying and amplifying DNA. The advantages of PCR will be highlighted by contrasting the technique with cloning, the components required and the stages of the process.
The author explains that Fabry disease is a genetic disorder that is inherited from parents to the progeny. It is caused by a mutation in alpha galactosidase gene that results in alpha galactosidase A enzyme deficiency. This is a X-linked gene carried on a mother’s X chromosome and the sons of a mother with this mutation will have 50%.
PCR has a close connection with the natural principle of DNA replication and it is associated with three step processes which are referred to as a cycle which repeats in a specific number of times. The important steps include denaturation, annealing and also extension.
Typically, the process involves a gene incorporation of a foreign gene to the target organism genome. The steps involved for creating genetically modified organism, using maize as an example are. 1. Isolation of the desired gene The very first step is identification of an existing organism exhibiting favourable traits; these traits are traced to the chromosomal characteristics of the known organism.
The Polymerase chain reaction was first introduced Kary Mullis in the 1980's (Bartlett et al 2003). Prior to its use in molecular biology, the amplification of DNA could only be carried out by cloning. PCR allows a "direct amplification without the use of cloning" (Bastianutto et al 2006).
The major constituents of PCR are, 1) DNA polymerase 2) short DNA fragment as primer 3) dNTPs 4) buffer and 5) DNA template. Invention of thermo stable DNA polymerase from thermophilic bacterium Thermus aquaticus (Taq Pol) (2) has made PCR a routine process in all molecular biology laboratories.
Presence of bands in both 100 bp and 400 bp position shows you are heterozygous showing polymorphism in one allele and normal gene in the other (+/- or long/short).
The 94C step in the procedure results in
The latter occurrence was the renamed Ebola River. Ebola assumes an incubation period of two 2 to 25 days before the onset of the symptoms. The prior symptoms consist of muscle pain, fever fatigue, sore throat and headache. These are closely followed by
The genetic factor is a vast molecule made by linking series of repeating units termed as nucleotides. Nucleotide has sugar phosphorus and a nitrogen called base. Bases with the DNA structure are adenine (A), guanine (G),
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