SacI restriction enzyme

Restriction Enzymes SacI

Experiment

Restriction Enzymes SacI

Product

SacI restriction enzyme from Takara Bio Inc

Manufacturer

Takara Bio Inc

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Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The 5′ PpAOS2 genomic fragment was digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) and inserted into pTN186, which contained a hygromycin-resistance cassette and had been digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) to obtain pTN186-PpAOS2KO5′. The 3′ PpAOS2 genomic fragment was inserted into pTN186-PpAOS2KO5′, which had been digested with SphI (Takara Bio Inc., Shiga, Japan) and SacI (Takara Bio Inc., Shiga, Japan), to yield pTN186-PpAOS2KO.

Protocol tips
The 5′ PpAOS2 genomic fragment was digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) and inserted into pTN186, which contained a hygromycin-resistance cassette and had been digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) to obtain pTN186-PpAOS2KO5′. The 3′ PpAOS2 genomic fragment was inserted into pTN186-PpAOS2KO5′, which had been digested with SphI (Takara Bio Inc., Shiga, Japan) and SacI (Takara Bio Inc., Shiga, Japan), to yield pTN186-PpAOS2KO.

Protocol tips

The 5′ PpAOS2 genomic fragment was digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) and inserted into pTN186, which contained a hygromycin-resistance cassette and had been digested with KpnI (Takara Bio Inc., Shiga, Japan) and HindIII (Takara Bio Inc., Shiga, Japan) to obtain pTN186-PpAOS2KO5′. The 3′ PpAOS2 genomic fragment was inserted into pTN186-PpAOS2KO5′, which had been digested with SphI (Takara Bio Inc., Shiga, Japan) and SacI (Takara Bio Inc., Shiga, Japan), to yield pTN186-PpAOS2KO.

Publication protocol

The PCR product was treated with SmaI and cloned into the SmaI site of pSTk14. The resultant plasmid, pSTk79, was linearized by Aor51HI digestion before integration at the leu1 locus.

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Manufacturer protocol

Download the product protocol from Takara Bio Inc for SacI restriction enzyme below.

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