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Micro RNAs (miRNAs) are short non-coding RNA oligonucleotides that negatively
regulate gene expression by incomplete base pairing with the untranslated (3UTR)
regions of the target mRNA. miRNAs implicates to a variety of basic cellular functions
et al, 2009). They are potential biomarkers in several human diseases and cancers.
(Bartels, C.L & Tsongalis, G.J, 2009). Several miRNAs have intriguing specific
expression as they have been identified in many different types of cancers including
pancreatic cancer, renal cancer, breast and colon cancer to determine the blood based
biomarkers. Alterations in the levels of miRNAs have been observed in several types of
cancer and high copy number variation appears to be a common cause and over
The urine derived miRNAs holds a gateway to explore the possible outcomes for early
prognosis for many potential diseases. Urine based biomarkers can be evaluated as non-
invasive indicators of kidney cancers. Exosomes are tiny vesicles are present abundantly
in urine and can be acquired my non-invasive methods and are enriched with miRNAs
and assists in robust assays that would be essential as predictive biomarkers of renal
The objective of this project is to optimize the best method of purification of exosomes
also, to examine if the yield of exosomes can be improved for the detection of miRNAs
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by performing different assays, also to effectively quantify the amount of exosomes by
each method. Most miRNAs are hallmark of several diseases, including cancer. Real-
time PCR was used to quantify the gene expression levels and the quality of RNA
achieved was detected by Agilent Bioanalyser. The purity of exosomes can also be
characterise the most effective method of isolation renders an avenue for investigating the
roles of proteins present within these vesicles. This study also involved the involvement
isolation. The possible elimination of this protein by the addition of Dithiothreitol (DTT)
is also studied. The direct involvement of miRNAs in causing the disease can be used for
identification of the certain biomarkers for the disease and in future, the results of this