Evaluation of Stem-Loop Reverse Transcription and Poly-A Tail Extension in MicroRNA Analysis of Body Fluids

Dunnett, Hannah, Williams, Graham and Van der Meer, Dieudonne (2015) Evaluation of Stem-Loop Reverse Transcription and Poly-A Tail Extension in MicroRNA Analysis of Body Fluids. MicroRNA, 4. ISSN 2211-5366

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Abstract

MicroRNA has been demonstrated to be a viable tool for body fluid identification purposes in forensic casework. Stem-loop reverse transcription (slRT) is regularly used for cDNA synthesis from mature miRNA, along with poly-A tail extension. Both have been used in a forensic context, but no direct comparison has been carried out. It has also not been shown whether poly-A tail extension can be used upon DNA extracts, as previously shown with slRT. Blood and saliva samples were collected and underwent DNA extraction with or without on-column DNA digestion. All samples were then aliquoted and underwent slRT and poly-A tail extension separately. qPCR was then conducted targeting microRNA markers hsa-miR-451 and hsa-miR-205. It was shown that the DNA digestion step did not affect the ability to differentiate between blood and saliva. It was also shown that this differentiation was possible using poly-A tail extension, and that poly-A tail extension exhibited more amplification than slRT. So whilst the choice of slRT and poly-A tail extension for the purpose of forensic body fluid identification is not critical, it may be best to use poly-A tail extension, particularly where there are low traces of sample.

Item Type:	Article
Subjects:	Q Science > Q Science (General) R Medicine > RA Public aspects of medicine > RA1001 Forensic Medicine. Medical jurisprudence. Legal medicine
Schools:	School of Applied Sciences School of Applied Sciences > Forensic Biology Research Group
Related URLs:	Author
Depositing User:	Cherry Edmunds
Date Deposited:	04 Feb 2015 12:24
Last Modified:	28 Aug 2021 18:32
URI:	http://eprints.hud.ac.uk/id/eprint/23261

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