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The beautiful people include Ben Aaron Johnsonn), a young botanist who grows terrific pot, and his pal and business partner Chon (Taylor Kitsch), a former Navy SEAL hero.
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For expression of GST-tagged MtbHadAB complex, cells were grown in Terrific Broth medium supplemented with 100 μg/mL ampicilin and 50 μg/mL kanamycin at 37°C.
Protein was overexpressed in E. coli BL21 DE3) CodonPlus-RIL cells (Stratagene) grown in terrific broth medium.
E. coli XL1-blue pREP cells were transformed with this construct and grown in Terrific Broth medium at 37°C until an OD600 of 1.
Briefly, saturated 40 mL overnight cultures grown in terrific broth (TB) plus 2% glucose at 30°C were transferred to 400 mL of fresh TB without glucose and shaken for 3 h at 25°C.
Cells were grown in Terrific Broth (1.2% peptone, 2.4% yeast extract, 0.94% K2HPO4 and 0.22% KH2PO4) (Difco; Sparks, MD) at 37°C to ∼0.8 OD600, at which time IPTG was added, the culture was cooled to 20°C in an ice bath, and the cells were incubated, shaking, for 12 hours.
The glycosyltransferases UGT71G1 and UGT73C8 from Medicago truncatula are excellent reagents for the regioselective glycosylation of (iso flavonoids in Escherichia coli grown in Terrific broth.
All KpClds were expressed in Escherichia coli Tuner (DE3) cells (Merck/Novagen) grown in Terrific Broth (TB) with ampicillin (100 μg/mL).
The pHIS8cyc1 and pHIS8cyc2 constructs were introduced into E. coli BL21 DE3) cells, and transformants were grown in terrific broth (TB) containing kanamycin (50 μg mL−1) at 37 °C.
E. coli BL21 (DE3) strains carrying these constructs were grown in Terrific broth (TB) or Luria-Bertani (LB) medium at 37°C until OD600 reached 0.7.
Cells were grown in terrific broth (Invitrogen, Grand Island, NY) to an OD595 of ∼0.9 and cold shocked for 20 min on ice.
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