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Restriction Fragment Pleomorphism - Coursework Example

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Restriction Fragment Pleomorphism

Restriction enzymes are highly specific in nature and single alteration in its recognize site (Restriction site) leads to complete inhibition of its activity. In RFLP technique particular gene having restriction site for an enzyme is used to generated DNA fragment(. If there was a mutation in this gene leads to altered restriction site which subsequently resultant in to DNA fragment having altered length. Based on size of a fragment and comparing with standard one can conclude mutation in given gene.
Here in case of sample A three bands were obtained when digested with DdeI indicating heterozygous for Gal6V while XhoI digest indicates homozygous for Gal6V clearly indicating anomaly in the results as digestion with two different enzymes give exactly opposite results. Comparing the banding pattern of Gel1A with theoretical banding of standard, there should be 3bands on XhoI digest compared to single band on DdI in case of Gal6V homozygous. But the banding pattern obtained on image 1A was not fitting in to the criteria indicating abnormal preparation. thus all the interpretation were drawn from gel 1B. in case of sample B there was three bands found corresponding to homozygous for CD5(-CT) standard sample while it was found to be heterozygous for GAL6V. Similarly sample C was found to be heterozygous for CD5(-CT) and heterozygous for Gal6V. Three control samples were given as standered namely C1 homozygus for Gal6V, C2- normal individual and C3 homozygus for CD5(-CT). Banding pattern obtained after digestion with DdeI and XhoI was as shown in fig 1A and 1B. Tabel 1 indicates phenotype and genotype of given sample.



Table 1: Summary of Genotype and Phenotype of given sample
Sample
DdeI enzyme
XhoI enzyme
Genotype
Phenotype
A
Glu6Val / +
CD5(-CT)/ CD5(-CT)
Heterozygous Glu6Val and
Homozygus CD5(-CT)
Thalassemia disease carrier of sickle cell anemia fetus
B
Glu6V/+
CD5(-CT)/ CD5(-CT)
Heterozygous Glu6Val and
Homozygus CD5(-CT)
Thalassemia disease carrier of sickle cell anemia fetus
C
Glu6Val / +
CD5(-CT)/ +
Heterozygous Glu6Val
Heterozygus CD5(-CT)
Carrier sickle cell
Crrier CD5(-CT)

Conclusion:
Genotyping of given sample were carried out using RFLP technique. The main objective of this study was to identify and differentiate closely related genetic disorder namely sickle cell anemia and thalassemia. Experimental results have indicated that there was some anomaly in electrophoretic patterns and thus experiments has to be performed again be for came in to any conclusion. Similarly experiments have demonstrated both aspect of RFLP at on side the technique was found to be easy, rapid and sensitive while other side it was found that it has some limitations like repeatability, requirement of large ...Show more

Summary

Genetic mutation or alteration in normal coding sequence leads to emergent of many life threatening diseases(Zhang 1994). the biggest challenges faced by physician an doctors are the incurable nature of these genetic diseases and thus it is highly recommended to identified high risk person and restrict transmission of these genetic mutation to offspring(Orkin SH 1984)…
Author : grahamkatherine
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