HpyF3I (DdeI) (10 U/µL)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	After amplification of the flaA fragment, a 10-μL sample (0.1–0.5 μg of amplified DNA quantified by Nanodrop®, Canada) of the amplicons was digested with the restriction enzyme HpyF3I (Thermo Scientific®) according to the manufacturer’s instructions. Briefly, 10 μL of PCR product, 18 μL nuclease-free water, 2 μL Buffer Tango (10×, Thermo Scientific®), and 1–2 μL of HpyF3I enzyme were added together in 0.2-mL DNase-free tubes and incubated at 37 °C for 16 h. Then, digests were analyzed on a 2.5% agarose gel containing 0.5 μg/mL ethidium bromide. Low-range DNA ladder (50 bp, Jena Bioscience) was used as standard for molecular size determination.

Protocol tips

After amplification of the flaA fragment, a 10-μL sample (0.1–0.5 μg of amplified DNA quantified by Nanodrop®, Canada) of the amplicons was digested with the restriction enzyme HpyF3I (Thermo Scientific®) according to the manufacturer’s instructions. Briefly, 10 μL of PCR product, 18 μL nuclease-free water, 2 μL Buffer Tango (10×, Thermo Scientific®), and 1–2 μL of HpyF3I enzyme were added together in 0.2-mL DNase-free tubes and incubated at 37 °C for 16 h. Then, digests were analyzed on a 2.5% agarose gel containing 0.5 μg/mL ethidium bromide. Low-range DNA ladder (50 bp, Jena Bioscience) was used as standard for molecular size determination.

Publication protocol

After amplification of the flaA fragment, a 10-μL sample (0.1–0.5 μg of amplified DNA quantified by Nanodrop®, Canada) of the amplicons was digested with the restriction enzyme HpyF3I (Thermo Scientific®) according to the manufacturer’s instructions. Briefly, 10 μL of PCR product, 18 μL nuclease-free water, 2 μL Buffer Tango (10×, Thermo Scientific®), and 1–2 μL of HpyF3I enzyme were added together in 0.2-mL DNase-free tubes and incubated at 37 °C for 16 h. Then, digests were analyzed on a 2.5% agarose gel containing 0.5 μg/mL ethidium bromide. Low-range DNA ladder (50 bp, Jena Bioscience) was used as standard for molecular size determination.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for HpyF3I (DdeI) (10 U/µL) below.

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