Gel Electophoresis and DNA Amplification - Essay Example

Amplifying the DNA prior to digestion is necessary because the process of amplification produces multiple copies of DNA sequences, thus increasing the amount of DNA required for the experiment. DNA amplification also increases the surface area for the action of the restriction enzyme because it focuses on a specific sequence only.

1. Controls for the experiment

The use of water in the amplification process was to make sure that the reagents are free of DNA contaminants.

The use of positive and negative controls in the whole process of the experiment can be necessary.  Positive controls should contain the same amount of components as those in the sample as well as a template that amplifies if the reaction proceeds according to plan. Positive control verifies the negative amplification results. Negative controls, specifically ‘no enzyme controls,’ are used to determine the origin of any amplification in the sample to be from the synthesized cDNA and not amplicon determinant or from genomic DNA. This is because the ‘no enzyme controls’ do not have reverse transcriptase.

2. Observations

This means that the gel electrophoresis will produce an extra line because of the extra fragment in the digested sample.

3. Distinguishing between normal individuals and patients with the mutation

Given that the amplification produces a DNA fragment of 450bp in length, normal individuals will only have a 445bp fragment while those with the mutation will have two fragments of 422 and 28. This means that the gel will show a single line in the normal sample and two lines in the patient sample.

4. Hematological tests

The first hematological test to be done is the activated protein C resistance test. This is done by testing blood’s resistance to activated protein C whereby the ratio of the number of clotting is determined with and without the activated protein C, an anti-clotting protein.

Genetic tests can also be done to determine if the individual inherited just one or two copies of the mutated gene.

5. Risks and treatment.

Patients with factor V Leiden and II 20210 mutations face the risk of initial venous thromboembolism or deep vein thrombosis and even pregnancy miscarriages.

Individuals who are carriers of the mutation and have experienced venous thromboembolism can be given heparin or oral warfarin to initiate anticoagulation and then continue with the treatment for 3-6 months after the episode of venous thromboembolism. Patients who have experienced episodes of venous thromboembolism or are at a high risk of occurrence of other episodes can undergo long-term anticoagulation therapy treatments.