Assessment of a fully automated high-throughput DNA extraction method from formalin-fixed,paraffin-embedded tissue for KRAS, and BRAF somatic mutation analysis |
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Authors: | Ronald van Eijk Lisa StevensHans Morreau Tom van Wezel |
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Institution: | Department of Pathology, Leiden University Medical Center, P.O.box 9600, L1-Q, Albinusdreef 2, 2300 RC Leiden, The Netherlands |
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Abstract: | Preoperative biopsies or imbedded cytological cells will become more and more a primary source of tissue for molecular diagnostic analyses as a result of novel neo-adjuvant treatment regimens for several cancer types. Furthermore there is a growing need to examine metastatic cancer tissue. Hence, nucleic acids need to be reliably isolated and analyzed from small amounts of formalin-fixed and paraffin-embedded (FFPE) tissue. The limited numbers of (tumor) cells in these samples make high quality and sensitive DNA isolation challenging. Also demands for faster turnaround times are growing. Therefore, we evaluated a fully automated DNA/RNA isolation system and compared this with a manual, classical routine molecular pathology method. We compared the quality of the isolates from both tissue cores and micro-dissection for detection of hotspot mutations in KRAS, BRAF applying hydrolysis probe assays. In addition we determined whether the automated method decreases the hands-on-time and turnaround times in routine molecular pathology workflow. |
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