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.
| Original language | English |
|---|---|
| Pages (from-to) | 150-154 |
| Number of pages | 5 |
| Journal | MicroRNA (Shāriqah, United Arab Emirates) |
| Volume | 3 |
| Issue number | 3 |
| Publication status | Published - 2014 |
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Evaluation of stem-loop reverse transcription and poly-A tail extension in microRNA analysis of body fluids. / Dunnett, Hannah; van der Meer, Dieudonne; Williams, Graham A.
In: MicroRNA (Shāriqah, United Arab Emirates), Vol. 3, No. 3, 2014, p. 150-154.Research output: Contribution to journal › Article
TY - JOUR
T1 - Evaluation of stem-loop reverse transcription and poly-A tail extension in microRNA analysis of body fluids
AU - Dunnett, Hannah
AU - van der Meer, Dieudonne
AU - Williams, Graham A.
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84991239260&partnerID=8YFLogxK
M3 - Article
VL - 3
SP - 150
EP - 154
JO - MicroRNA (Shariqah, United Arab Emirates)
JF - MicroRNA (Shariqah, United Arab Emirates)
SN - 2211-5366
IS - 3
ER -