Restriction Fragment Pleomorphism Coursework Example | Topics and Well Written Essays - 1000 words

breastwork Fragment Pleomorphism - Coursework ExampleRestriction enzymes are highly specific in nature and single alteration in its recognize land 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 deoxyribonucleic acid fragment(. If there was a mutation in this gene leads to altered restriction site which subsequently consecutive in to DNA fragment having altered length. Based on size of a fragment and comparison with standard one can conclude mutation in given gene.Here in racing shell of sample A trine 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 hypothetic banding of standard, there should be 3bands on XhoI digest compared to s ingle band on DdI in fact 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. alike 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 genetic constitution and Phenotype of given sampleSampleDdeI enzymeXhoI enzymeGenotypePhenotypeAGlu6Val / +CD5(-CT)/ CD5(-CT)heterozygous Glu6Val and Homozygus CD5(-CT)Thalassemia disease carrier of sickle booth anemia fetus BGlu6V/+CD5(-CT)/ CD5 (-CT)Heterozygous Glu6Val and Homozygus CD5(-CT)Thalassemia disease carrier of sickle cell anemia fetusCGlu6Val / +CD5(-CT)/ +Heterozygous Glu6ValHeterozygus CD5(-CT)Carrier sickle cellCrrier CD5(-CT)Conclusion Genotyping of given sample were carried out using RFLP technique. The main nonsubjective 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 military position 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