BL21-pG-KJE10

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The TOP10 (Invitrogen™), BL21-CodonPlus (DE3)-RIL (Agilent) and BL21-pG-KJE8 (Takara Bio Inc) strains were transformed with the recombinant plasmids. For expression assays, a positive colony for each construct in each strain was grown in LB medium that was supplemented with 100 μg/ml ampicillin and 50 μg/ml chloramphenicol overnight at 37° C with constant stirring at 200 rpm.	The following day, each culture was diluted with fresh medium, and for the BL21-pG-KJE8 system, the medium was supplemented with 1 mg/ml arabinose and 10 ng/ml tetracycline (chaperone expression-inducing agents). The cultures were incubated until they reached an optical density (OD600) between 0.6 and 0.8, time at which isopropyl thio-beta-galactopyranoside (IPTG, 0.1 mM) was added, and growth was allowed for 16 h at 37° C.

Protocol tips
The TOP10 (Invitrogen™), BL21-CodonPlus (DE3)-RIL (Agilent) and BL21-pG-KJE8 (Takara Bio Inc) strains were transformed with the recombinant plasmids. For expression assays, a positive colony for each construct in each strain was grown in LB medium that was supplemented with 100 μg/ml ampicillin and 50 μg/ml chloramphenicol overnight at 37° C with constant stirring at 200 rpm.
Downstream tips
The following day, each culture was diluted with fresh medium, and for the BL21-pG-KJE8 system, the medium was supplemented with 1 mg/ml arabinose and 10 ng/ml tetracycline (chaperone expression-inducing agents). The cultures were incubated until they reached an optical density (OD600) between 0.6 and 0.8, time at which isopropyl thio-beta-galactopyranoside (IPTG, 0.1 mM) was added, and growth was allowed for 16 h at 37° C.

Publication protocol

"The TOP10 (Invitrogen™), BL21-CodonPlus (DE3)-RIL (Agilent) and BL21-pG-KJE8 (Takara Bio Inc) strains were transformed with the recombinant plasmids. For expression assays, a positive colony for each construct in each strain was grown in LB medium that was supplemented with 100 μg/ml ampicillin and 50 μg/ml chloramphenicol overnight at 37° C with constant stirring at 200 rpm.
The following day, each culture was diluted with fresh medium, and for the BL21-pG-KJE8 system, the medium was supplemented with 1 mg/ml arabinose and 10 ng/ml tetracycline (chaperone expression-inducing agents). The cultures were incubated until they reached an optical density (OD600) between 0.6 and 0.8, time at which isopropyl thio-beta-galactopyranoside (IPTG, 0.1 mM) was added, and growth was allowed for 16 h at 37° C. "

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Papers

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Manufacturer protocol

Download the product protocol from Jacqueline Chaparro-Olaya, Laboratorio de Parasitología Molecul for BL21-pG-KJE10 below.

Videos

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