دورية أكاديمية
أعرض في EDS

Effects of storage conditions on the stability of qPCR reagents: implications for environmental DNA detection.

التفاصيل البيبلوغرافية
العنوان: Effects of storage conditions on the stability of qPCR reagents: implications for environmental DNA detection.
المؤلفون: Lopez MLD; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada., Crichton EM; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada.; Pacific Biological Station, Fisheries and Oceans Canada, Nanaimo, BC, V9T 6N7, Canada., Allison MJ; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada., Dema AH; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada., Bonderud MT; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada., Helbing CC; Department of Biochemistry & Microbiology, University of Victoria, Victoria, BC, V8P 5C2, Canada. chelbing@uvic.ca.
المصدر: BMC research notes [BMC Res Notes] 2024 Jul 18; Vol. 17 (1), pp. 199. Date of Electronic Publication: 2024 Jul 18.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Biomed Central Country of Publication: England NLM ID: 101462768 Publication Model: Electronic Cited Medium: Internet ISSN: 1756-0500 (Electronic) Linking ISSN: 17560500 NLM ISO Abbreviation: BMC Res Notes Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London : Biomed Central, 2008.
مواضيع طبية MeSH: Real-Time Polymerase Chain Reaction*/methods , Real-Time Polymerase Chain Reaction*/standards , DNA, Environmental*/analysis , DNA, Environmental*/genetics, Indicators and Reagents ; Freezing ; DNA Primers/genetics ; Specimen Handling/methods
مستخلص: Objective: Environmental DNA (eDNA) detection is a transformative tool for ecological surveys which in many cases offers greater accuracy and cost-effectiveness for tracking low-density, cryptic species compared to conventional methods. For the use of targeted quantitative PCR (qPCR)-based eDNA detection, protocols typically require freshly prepared reagents for each sample, necessitating systematic evaluation of reagent stability within the functional context of eDNA standard curve preparation and environmental sample evaluation. Herein, we assessed the effects of long-term storage and freeze-thaw cycles on qPCR reagents for eDNA analysis across six assays.
Results: Results demonstrate qPCR plates (containing pre-made PCR mix, primer-probe, and DNA template) remain stable at 4 °C for three days before thermocycling without fidelity loss irrespective of qPCR assay used. Primer-probe mixes remain stable for five months of - 20 °C storage with monthly freeze-thaw cycles also irrespective of qPCR assay used. Synthetic DNA stocks maintain consistency in standard curves and sensitivity for three months under the same conditions. These findings enhance our comprehension of qPCR reagent stability, facilitating streamlined eDNA workflows by minimizing repetitive reagent preparations.
(© 2024. The Author(s).)
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معلومات مُعتمدة: PDF Fellowship Liber Ero Foundation; 312ITD Genome Canada; 312ITD Genome British Columbia; 312ITD Genome Quebec
فهرسة مساهمة: Keywords: Freeze–thaw cycles; Primer and probe stability; Synthetic DNA; eDNA; qPCR master mix
المشرفين على المادة: 0 (DNA, Environmental)
0 (Indicators and Reagents)
0 (DNA Primers)
تواريخ الأحداث: Date Created: 20240718 Date Completed: 20240719 Latest Revision: 20240725
رمز التحديث: 20240726
مُعرف محوري في PubMed: PMC11264737
DOI: 10.1186/s13104-024-06850-4
PMID: 39026307
قاعدة البيانات: MEDLINE
الوصف
تدمد:1756-0500
DOI:10.1186/s13104-024-06850-4