PV92 practical - Assignment Example

This would, therefore, mean that the genomic DNA is not protected from degradation by the cellular enzymes released during the isolation protocol of the DNA. Response to Question 2. In order to carry out a PCR, one requires DNA. This means a cell is important since it is the source of the DNA. The cell can also serve as a source for base pairs, enzymes and primers. Response to Question 3. There are various structures that are often broken in order to release the DNA from a cell. These include the cells, the cheek tissues, the phosholids, the nucleus, and proteinase K.

this is done so that proteins along with other organically-soluble compounds are removed from chromosomes and cells. PCR Amplication. Response to question 1. It is important to have a primer on either side of the segment of a DNA that is going to be amplified because these form the four basic bases. Uracil exists in RNA, despite being a special case. Primers are vital and essential components of the synthesis. They synthesize an RNA short piece which is complementary to the strant of the template DNA forming the bonds of hydrogen with it.

This provides a DNA polymer with a starting poing which that is required for the synthesis. After the completion of the DNA synthesis, the segment of RNA is removed and a DNA replaces it. Response to Question 2. Taq DNA polymerase is the thermostable DNA polymerase that derived its name from the thermophilic bacterium called Thermus aquaticus. The T.aquaticus is a specific bacterium found in hydrothermal and hot springs vents. Response to Question 3. It is because they are the only four basic bases with uracil being is present in the RNA, although it is a special case.

Components of Master mix include water, oug template, the buffer, deoxynucleotides, aum primers, template DNA and ampliTaq polymerase. The buffer is used for keeping the master mix at a suitable PH in order to make sure the reaction of the PCR occurs. Deoxynu-cleotides, in this case help in providing the energy and the nucleosides used for synthesizing the DNA. As widely cited, it is vital adding equal amounts of each of the nucleotide (, dTTP, dCTP, dATP and dGTP) onto the master matrix for purposes of preventing the mismatch of bases.

The ouM primers are short pieces of the DNA that often bind onto the DNA template to allow the Taq DNA polymerase enzyme to start the incorporation of deoxynucleotides. The AmpliTaq polymerase is used, in this case, to add the deoxynucleotides onto the DNA template. Last but not least, there is presence of oug template that is often amplified by a PCR reaction. Response to Question 4. The DNA PCR amplification takes place in repeated cycles of the three temperature-dependent steps. In the first step, a double-stranded DNA or (dsDNA) template becomes denatured.

In the second stage, the oligonucleotide primers become annealed to a single-stranded DNA also called (ssDNA) template. In this case one primer become designed specifically to anneal onto a certain specific region found on the left side of the strands of the DNA with the other primer being designed specifically to anneal onto a specific region of the complementary DNA strand right side. In the third stage, the DNA polymerase is meant to extend on each primer in 3' to 5' direction, while duplicating the fragment of the DNA between the primers. While