Vibration as a pitfall in pyrosequencing analyses
| Autor: | Hannah Sturm, Thomas Bajanowski, Laura Schäfer, Helen Konrad, Micaela Poetsch, Lena Hördt |
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| Rok vydání: | 2021 |
| Předmět: |
Computer science
Medizin High-Throughput Nucleotide Sequencing Reproducibility of Results Sequence Analysis DNA DNA Methylation Vibration Purchasing Pathology and Forensic Medicine Reliability engineering Methylation analysis Pyrosequencing Humans CpG Islands Reliability (statistics) Forensic genetics |
| Zdroj: | International journal of legal medicine. 136(1) |
| ISSN: | 1437-1596 |
| Popis: | Since methylation analysis has become an important tool in forensic genetics, the reliability and credibility of the method must be ensured. After a successful validation and establishment of several pyrosequencing assays using a PyroMark® Q48 Autoprep instrument (Qiagen, Hilden, Germany), we decided to expand the method further purchasing a second instrument. But after initializing this second instrument side by side with the first, the majority of analyses failed (97 samples of 133 samples (73%)). The number of error messages increased rapidly and the average RFU values decreased. After purchasing two anti-vibration weighing tables for the PyroMark® instruments and repeating the analyses under the same conditions and with identical samples the results improved considerably, 115 samples of 130 samples (88%) showed successful and reproducible results. These findings demonstrate the impact of vibrations and percussions on PyroMark® Q48 Autoprep performance and the reliability of methylation analyses. |
| Databáze: | OpenAIRE |
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