Paul.xu: Created page with "== BL21 AI Expresion of toxic human proteins
==
1. Transform BL21 AI (Invitrogen) with pRARE2 plasmid. Plate on LB/Chl plate.
2. Transform BL21 AI/pRare2 with exp..."

2013-02-14T20:22:50Z

Created page with "== BL21 AI Expresion of toxic human proteins == 1. Transform BL21 AI (Invitrogen) with pRARE2 plasmid. Plate on LB/Chl plate. 2. Transform BL21 AI/pRare2 with exp..."

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== BL21 AI Expresion of toxic human proteins ==

 

1. Transform BL21 AI (Invitrogen) with pRARE2 plasmid. Plate on LB/Chl plate. 

2. Transform BL21 AI/pRare2 with expression plasmid . 

3. Grow in 1 L of TB media (12 g tryptone, 24 g yeast extract, 2.31 g KH2 PO4 (mono-basic), 12.5 g K 2 HPO4 (dibasic), 40 mL glycerol) containing 35 μg∕mL chloramphenicol and 50 μg∕mL kanamycin or 100  μg∕mL Amp at 37 °C until OD600 1⁄4 1.0.

4. Cool culture to 20 °C.

5. Add 4 mL of 50% w∕v arabinose and 1 M isopropyl-β-D-thiogalactopyranose (IPTG) to a final concentration of 0.4 mM.

6. Continue to culture for 1 hr at 20 °C.

7. Incubate the culture o/n at 20 oC.

8. Harvest cells by centrifugation.

 

[[Category:Protocols]]

BL21-AI - Revision history

Paul.xu: Created page with "== BL21 AI Expresion of toxic human proteins == 1. Transform BL21 AI (Invitrogen) with pRARE2 plasmid. Plate on LB/Chl plate. 2. Transform BL21 AI/pRare2 with exp..."

Paul.xu: Created page with "== BL21 AI Expresion of toxic human proteins
==
1. Transform BL21 AI (Invitrogen) with pRARE2 plasmid. Plate on LB/Chl plate.
2. Transform BL21 AI/pRare2 with exp..."