PvuI restriction enzyme

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Finally, the resulting pEX12–tTA2S was digested with NotI and PvuI Takara) to obtain a 7,627 bp fragment (PmThy1.2–tTA2S–p(A)) for pronuclear injection NotI and PvuI (Takara) to obtain a 7,627 bp fragment (PmThy1.2–tTA2S–p(A)) for pronuclear injection (Fig. 1a).

Protocol tips
Finally, the resulting pEX12–tTA2S was digested with NotI and PvuI Takara) to obtain a 7,627 bp fragment (PmThy1.2–tTA2S–p(A)) for pronuclear injection NotI and PvuI (Takara) to obtain a 7,627 bp fragment (PmThy1.2–tTA2S–p(A)) for pronuclear injection (Fig. 1a).

Publication protocol

"In order to restrict the expression of the tTA2 transactivator to the brain, we first excised the TA2S-encoding cDNA from the pTet-Off Advanced Vector (Cat. No. 631070—Clontech) with EcoRI and
BamHI (both Takara), and performed subsequent Klenow (Takara) filling of protruding ends. A pEX12 containing the partially recombinant mouse neuronal
specific mThy1.2 promoter (Ittner et al. 2008) was linearized with XhoI (Takara), blunt ended with Klenow and dephosphorylated. The tTA2S fragment
was ligated into pEX12 using T4 ligase (Promega). Finally, the resulting pEX12–tTA2S was digested with NotI and PvuI Takara) to obtain a 7,627 bp fragment (PmThy1.2–tTA2S–p(A)) for pronuclear injection
NotI and PvuI (Takara) to obtain a 7,627 bp fragment
(PmThy1.2–tTA2S
–p(A)) for pronuclear injection
(Fig. 1a)."

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