FastDigest Tru1I

Protocol tips

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	The nested PCR product was digested with TaaI and Tru1I FastDigest enzymes (Thermo Fisher Scientific Inc., Waltham, MA, USA) that produce diagnostic band patterns in agarose gel allowing to identify the two targeted species. With TaaI enzyme, brown trout DNA gives two bands of 205 and 272 bp, and with Tru1I, rainbow trout DNA gives a band of 66 bp.

Protocol tips
The nested PCR product was digested with TaaI and Tru1I FastDigest enzymes (Thermo Fisher Scientific Inc., Waltham, MA, USA) that produce diagnostic band patterns in agarose gel allowing to identify the two targeted species. With TaaI enzyme, brown trout DNA gives two bands of 205 and 272 bp, and with Tru1I, rainbow trout DNA gives a band of 66 bp.

Publication protocol

"Species-specific PCR-RFLP
The nested PCR-RFLP method was developed as described in Clusa et al. (2017) for
detecting the two targeted species (brown and rainbow trout). Briefly: a first PCR was
carried out to amplify a 567 bp fragment of the 16S rRNA gene, using as forward the 16Snew-F primer designed in the cited research (Table 3), and the reverse 16S-Br universal
primer from Palumbi et al. (2002). A nested PCR amplification was then performed with
the pair of Salmonidae-specific primers described in Zaiko et al. (2015) (Table 3). The
nested PCR product was digested with TaaI and Tru1I FastDigest enzymes (Thermo Fisher
Scientific Inc., Waltham, MA, USA) that produce diagnostic band patterns in agarose gel
allowing to identify the two targeted species. With TaaI enzyme, brown trout DNA gives
two bands of 205 and 272 bp, and with Tru1I, rainbow trout DNA gives a band of 66 bp"

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Manufacturer protocol

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