e-journal

Recognizing false positives: synthetic oligonucleotide controls for environmental DNA surveillance

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1. EnvironmentalDNA(eDNA) is increasingly used for surveillance and detection of species of interest in aquatic
and soil samples.
2. A significant risk associated with eDNAmethods is potential false-positive results due to laboratory contamination.
3. To minimize and quantify this risk, we designed and validated a set of synthetic oligonucleotides for use as
species-specific positive PCR controls for several high-profile aquatic invasive species.
4. The controls consist of species-specific sequences for the species of interest, with the addition of a synthetic
insert containing recognition sites for several restriction enzymes.
5. Following PCR, the presence of the synthetic insert can be detected using gel electrophoresis, restriction
enzyme digests orDNAsequencing. For quantitative PCR (qPCR), false positives in environmental samples can
also be detected using a fluorescent probe designed to detect the synthetic insert.
6. The generation of synthetic controls is a cost-effective, reproducible method that increases the power and reliability
of eDNAtesting by eliminatingmisinterpretation of false-positive results from laboratory contamination.

Key-words: contamination, (eDNA), environmental DNA, false positives, synthetic controls

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Ketersediaan

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Informasi Detail

Judul Seri

Methods in Ecology and Evolution

No. Panggil

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Penerbit

: British Ecological Society., 2016

Deskripsi Fisik

Methods in Ecology and Evolution 2016, 7, 23–29

Bahasa

English

ISBN/ISSN

doi: 10.1111/2041-21

Klasifikasi

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Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

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Subjek

BIOLOGI

Info Detail Spesifik

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Pernyataan Tanggungjawab

Wati/Agus

Versi lain/terkait

Lampiran Berkas

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