Evaluation of stem-loop reverse transcription and poly-A tail extension in microRNA analysis of body fluids

WILLIAMS, Graham, Dunnett, Hannah and van der Meer, Dieudonne (2014) Evaluation of stem-loop reverse transcription and poly-A tail extension in microRNA analysis of body fluids. MicroRNA, 3 (3). pp. 150-154. ISSN 2211-5374

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Abstract or description

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
Faculty:	School of Law, Policing and Forensics > Criminal Justice and Forensic Science
Depositing User:	Graham WILLIAMS
Date Deposited:	30 Aug 2017 15:14
Last Modified:	24 Feb 2023 13:48
URI:	https://eprints.staffs.ac.uk/id/eprint/3760

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