Comparative evaluation of different extraction and quantification methods for forensic RNA analysis

Grabmüller, Melanie; Madea, Burkhard; Courts, Cornelius

doi:10.1016/j.fsigen.2015.01.006

Cited by 39 publications

(25 citation statements)

References 73 publications

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“…Evaluating all extraction procedures at the same time, saliva samples show a higher miRNA yield than blood samples (blood: 264.2 pg/μL; saliva: 419.35 pg/μL) which is in concordance with Grabmüller et al. . Surprisingly, the extraction efficiency was higher for blood samples (blood: 76.5%; saliva: 59 %).…”

Section: Resultssupporting

confidence: 87%

“…The extraction procedure is a very influential step for the success of the subsequent analyses steps and needs to be as efficient as possible. Some methods for co‐isolation of miRNA and DNA from blood and saliva have already been tested [, ] but those studies were not able to present a procedure suitable for routine casework.…”

Section: Resultsmentioning

confidence: 99%

“…Also, the miRNA extraction efficiency was the highest for the automated extraction procedure for both body fluids (blood ( Fig. 2F [32]. Surprisingly, the extraction efficiency was higher for blood samples (blood: 76.5%; saliva: 59 %).…”

Section: Mirna Quantification and Expression Analysismentioning

confidence: 93%

“…According to Grabmüller et al. , the NucleoSpin® miRNA Kit (Macherey Nagel, Düren) showed the best performance for DNA and miRNA purification.…”

Section: Methodsmentioning

confidence: 99%

“…According to Grabmüller et al [32], the NucleoSpin R miRNA Kit (Macherey Nagel, Düren) showed the best performance for DNA and miRNA purification. Actually, this kit was designed to purify miRNA, so DNA extraction was done from the swabs, which were reused after the incubation for miRNA extraction applying the Maxwell R 16 LEV Blood DNA Kit.…”

Section: Semi-manual Extraction Procedure-procedures 2 (P2)mentioning

confidence: 99%

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Automation of DNA and miRNA co‐extraction for miRNA‐based identification of human body fluids and tissues

et al. 2016

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In the last years, microRNA (miRNA) analysis came into focus in the field of forensic genetics. Yet, no standardized and recommendable protocols for co-isolation of miRNA and DNA from forensic relevant samples have been developed so far. Hence, this study evaluated the performance of an automated Maxwell® 16 System-based strategy (Promega) for co-extraction of DNA and miRNA from forensically relevant (blood and saliva) samples compared to (semi-)manual extraction methods. Three procedures were compared on the basis of recovered quantity of DNA and miRNA (as determined by real-time PCR and Bioanalyzer), miRNA profiling (shown by Cq values and extraction efficiency), STR profiles, duration, contamination risk and handling. All in all, the results highlight that the automated co-extraction procedure yielded the highest miRNA and DNA amounts from saliva and blood samples compared to both (semi-)manual protocols. Also, for aged and genuine samples of forensically relevant traces the miRNA and DNA yields were sufficient for subsequent downstream analysis. Furthermore, the strategy allows miRNA extraction only in cases where it is relevant to obtain additional information about the sample type. Besides, this system enables flexible sample throughput and labor-saving sample processing with reduced risk of cross-contamination.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Mirna Quantification and Expression Analysismentioning

confidence: 93%

“…According to Grabmüller et al. , the NucleoSpin® miRNA Kit (Macherey Nagel, Düren) showed the best performance for DNA and miRNA purification.…”

Section: Methodsmentioning

confidence: 99%

Section: Semi-manual Extraction Procedure-procedures 2 (P2)mentioning

confidence: 99%

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Automation of DNA and miRNA co‐extraction for miRNA‐based identification of human body fluids and tissues

et al. 2016

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Autopsy

Knudsen¹,

Thomsen²,

Grellner³

et al. 2022

Handbook of Forensic Medicine

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Isolation of enriched small RNA from cell‐lysate using on‐chip isotachophoresis

2019

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In spite of the growing interest in the roles and applications of small RNAs (sRNAs), sRNA isolation methods are inconsistent, tedious, and dependent on the starting number of cells. In this work, we employ ITP to isolate sRNAs from the cell‐lysate of K562 (chronic myelogenous leukemia) cells in a polydimethylsiloxane (PDMS) mesofluidic device. Our method specifically purifies sRNA of <60 nucleotides from lysate of a wide range of cell number spanning from 100 to 1 000 000 cells. We measured the amount of sRNA using the Agilent Bioanalyzer and further verified the extraction efficiency by reverse transcription quantitative PCR. Our method was shown to be more efficient in sRNA extraction than commercial sRNA isolation kits, especially when using smaller numbers of starting cells. Our assay presents a simple and rapid sRNA extraction method with 20 min assay time and no intermediate transfer steps.

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Comparative evaluation of different extraction and quantification methods for forensic RNA analysis

Cited by 39 publications

References 73 publications

Automation of DNA and miRNA co‐extraction for miRNA‐based identification of human body fluids and tissues

Automation of DNA and miRNA co‐extraction for miRNA‐based identification of human body fluids and tissues

Autopsy

Isolation of enriched small RNA from cell‐lysate using on‐chip isotachophoresis

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