Influence of Cold Ischemia Time and Storage Period on DNA Quality and Biomarker Research in Biobanked Colorectal Cancer Tissues
| العنوان: | Influence of Cold Ischemia Time and Storage Period on DNA Quality and Biomarker Research in Biobanked Colorectal Cancer Tissues |
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| المؤلفون: | Roh Mee-Sook, Pak Min-Gyoung |
| المصدر: | Kosin Medical Journal. 35:26-37 |
| بيانات النشر: | Kosin University College of Medicine, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Oncology, medicine.medical_specialty, Colorectal cancer, business.industry, Period (gene), Ischemia, medicine.disease, Cold Ischemia Time, chemistry.chemical_compound, chemistry, Internal medicine, medicine, Biomarker (medicine), business, DNA |
| الوصف: | Objectives Biobanking plays an important role in future research. Assessment and control of the preanalytical variables of biobanked tissues are fundamentals for the optimal use of biospecimens. Methods Forty-five colorectal cancer (CRC) tissues stored at −80°C in Bio-Resource Bank were evaluated to define the influence of cold ischemia time (CIT) and storage period (SP) on DNA quality in biobanked tissues. Three CITs (less than 30 minutes (CIT-1), 30–45 minutes (CIT-2), and 45–60 minutes (CIT-3)) and three SPs (less than 1 year (SP-1), 2–3 years (SP-2), and 4–5 years (SP-3)) were chosen. NanoDrop spectrophotometer was used to determine the 260/280 ratio for DNA purity. DNA integrity was analyzed by a UV transilluminator following electrophoresis on 2% agarose gel. To evaluate the practical usability of DNA for biomarker research, KRAS mutation status was assessed by PCR amplification. Results All DNA specimens had a 260/280 ratio ranging between 1.8 and 2.0 with the exception of one specimen (CIT-2/SP-2 group). For DNA integrity, DNA appeared as a compact, high-molecular-weight band with no or scanty low-molecular-weight smears. The concordance of KRAS mutation status between paired biobanked frozen tissues and formalin-fixed paraffin-embedded tissues was 100%. DNA remained stable in CRC tissues kept at room temperature for up to 1 hour and long-term storage up to 5 years. Conclusions Storage conditions of our biobank are suitable for long-term (at least five years) specimen preservation with high DNA quality. These results have practical implications that could affect banking guidelines. |
| تدمد: | 2586-7024 2005-9531 |
| DOI: | 10.7180/kmj.2020.35.1.26 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::f1c22640e3ee26c09dbc0ad27c758b90 https://doi.org/10.7180/kmj.2020.35.1.26 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi...........f1c22640e3ee26c09dbc0ad27c758b90 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 25867024 20059531 |
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| DOI: | 10.7180/kmj.2020.35.1.26 |