PauI (BssHII) (10 U/µL)

Protocol tips

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	These plasmids were digested with ClaI and PauI (Thermo Fisher Scientific) and the Pbp1 polyglutamine sequence was gel purified and ligated into the full length Pbp1+ promoter plasmid. All plasmids had sequence fidelity confirmed by Sanger sequencing.

Protocol tips
These plasmids were digested with ClaI and PauI (Thermo Fisher Scientific) and the Pbp1 polyglutamine sequence was gel purified and ligated into the full length Pbp1+ promoter plasmid. All plasmids had sequence fidelity confirmed by Sanger sequencing.

Publication protocol

The vector pRS303 was used as a backbone for cloning. First, a TAP tag was cloned into the vector using the restriction enzymes PdiI and XhoI (Thermo Fisher Scientific). Next, a full length Pbp1 including the native promoter (460 bp upstream of Pbp1 start site) PCR product was generated from W303 genomic DNA using Phusion (NEB) and cloned into the TAP-tagged vector using PdiI and PauI (Thermo Fisher Scientific). To generate the polyglutamine plasmids, Pbp1 polyglutamine sequence plasmids (460 bp upstream to ClaI cut site within Pbp1 downstream of polyglutamine site) were ordered from Thermo Fisher Scientific. These plasmids were digested with ClaI and PauI (Thermo Fisher Scientific) and the Pbp1 polyglutamine sequence was gel purified and ligated into the full length Pbp1+ promoter plasmid. All plasmids had sequence fidelity confirmed by Sanger sequencing.

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Manufacturer protocol

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

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