NotI restriction enzyme

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	The amplicons of Tat-NGB-His and His-NGBHis, and the pPIC9K plasmid (prepared in our laboratory) were digested with restriction endonuclease EcoRI and NotI (TaKaRa, Japan), and ligated with T4 ligase (TaKaRa, Japan). Thus, the conjugated plasmids were transformed into E. coli Top10 competent cells (Invitrogen, USA) and identified by double digestion of restriction endonuclease EcoRI and NotI, and then direct sequencing.

Protocol tips

The amplicons of Tat-NGB-His and His-NGBHis, and the pPIC9K plasmid (prepared in our laboratory) were digested with restriction endonuclease EcoRI and NotI (TaKaRa, Japan), and ligated with T4 ligase (TaKaRa, Japan). Thus, the conjugated plasmids were transformed into E. coli Top10 competent cells (Invitrogen, USA) and identified by double digestion of restriction endonuclease EcoRI and NotI, and then direct sequencing.

Publication protocol

"The Tat-NGB-His and His-NGB-His cDNA fragments
were amplified, respectively, from pET28b(?)-NGB plasmids (prepared in our laboratory) by polymerase chain
reaction (PCR) using following primers as shown in
Table 1. The amplicons of Tat-NGB-His and His-NGBHis, and the pPIC9K plasmid (prepared in our laboratory)
were digested with restriction endonuclease EcoRI and
NotI (TaKaRa, Japan), and ligated with T4 ligase (TaKaRa,
Japan). Thus, the conjugated plasmids were transformed
into E. coli Top10 competent cells (Invitrogen, USA) and
identified by double digestion of restriction endonuclease
EcoRI and NotI, and then direct sequencing"

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