Bacto tryptone

Bacto-tryptone (1 g), bacto-yeast extract (0.5 g) and NaCl (1 g) were dissolved in 95 mL of deionized water and the pH was adjusted to 7.0 with 5 m NaOH. The volume was adjusted to 100 mL with deionized water. A portion of the resulting solution (5 mL) was placed in a 20 mL test tube with a poromeric silicone plug and the rest was placed in a 500 mL Sakaguchi flask with a poromeric silicone plug. The media were sterilized by autoclaving (121 °C, 20 min). 

Materials for media (Bacto Tryptone, Bacto Yeast Extract and Difco SOB Medium, agar powder and NaCl) and antibiotics (ampicillin sodium and kanamycin sulfate). 

Luria-Bertani (LB) medium. Each liter contains 10 g Bacto-tryptone, 5 g Bactoyeast extract, and 10 g NaCl. Adjust to pH 7.5 with NaOH. Ampicillin. A solution of the sodium salt is prepared in water (25mg/mL) and sterilized by passage through a microfilter (e.g., a 0.22 H Millipore filter). Store the solution in a freezer. 

LB broth 10 g/L Bacto-tryptone, 5 g/L Bacto-yeast extract, 10 g/L NaCl, adjusted to pH 7.0 with NaOH and sterilized by autoclaving. 

SB medium (SB) 30 g/L Bacto-tryptone (Difco, Detroit, MI), 20 g/L yeast extract (Difco), 10 g/L MOPS (3-[A/-morpholino]propanesulfonic acid), pH 7 0, containing 10 pg/mL tetracycline (Boehringer Mannheim, Indianapolis, IN)... 

TYE medium. 10 g Bacto-tryptone, 5 g yeast extract, 8 g NaCl made up to 1 L. Adjust pH to 7 4 with lMNaOH. 

SOC medium 2% (w/v) bacto tryptone, 0.5% (w/v) yeast extract, 10 mM NaCl, 2.5 mM KC1. After autoclaving add the following sterilized solutions at the final concentration indicated 10mMMgCl2, lOmM MgS04, 20mM glucose. 

Pellet cells for 5 min in two 50 mL Falcon tubes (e.g., 2000 rpm in a Beckman J-6M swinging bucket centrifuge with Beckman JS-4.2 rotor). (We use sterile Falcon tubes (BD Biosciences, San Jose, CA, cat. 352070) to minimize the chance of nuclease contamination.) The supernatant is removed. Optionally, one pellet can be resuspended in equal volume of LB (1% Bacto Tryptone, 0.5% yeast extract and 0.5% NaCl in water) and 60% glycerol, divided into multiple fractions and stored at —80 °C for future use. The other pellet is resuspended in 25 mL at 37 °C LB, then add into 2 L LB containing 2 mL of 50 mg/mL Carbenicillin and 2 mL of 10 mg/mL Chloramphenicol. Grow at 37 °C until the ODgoo is close to 1,... 

SOB medium 20g/L bacto-tryptone (Difco Laboratories), 5g/L bacto-yeast extract (Difco Laboratories), 0.5 g/L NaCl, 16.6 g/L KC1. 

Luria broth—Luria broth Prepare five 100-ml and five 10-liter solutions containing 10 g/liter Bacto-Tryptone, 5 g/liter Bacto-Yeast extract, 5 g/liter NaCl, 0.5 g/liter antifoam agent (e.g., Polyglycol P-2000), and 1 g/liter glucose (autoclave separately). Autoclave the main solution and separate solution of concentrated glucose for 20 min at 105 to 110°C and cool use sterile conditions to add the glucose solution to the main solution and use within 24 hr. 

X TY medium dissolve 20g/l Bacto-Tryptone (Difco), 10 g yeast extract (Difco), and 10 g NaCI in 1 I water, and autoclave... 

Bacto Tryptone Bacto Yeast Extract SP-Sepharose fast flow... 

YT media (10 g bacto-yeast extract, 16 g bacto-tryptone, 5 g NaCl water added to 1 L and pH adjusted to 7.0 with NaOH autoclaved). 

LB (5 g bacto-yeast extract, 10g bacto-tryptone, 10g NaCl water added to 1L autoclaved). 

SOC media (5g bacto-yeast extract, 20g bacto-tryptone,... 

Terrific Broth (TB) To 700 mL H2O, add 12 g Bacto Tryptone, 24 g yeast extract, 10 mL glycerol, and 2 g of glucose (tee Note 3). Stir to dissolve solutes then bring volume to 900 mL. Sterilize by autoclaving. Cool media to room temperature then add 100 mL of filter sterilized lOx phosphate buffer (100 mL 2.31 g KH2PO4,12.54 g K2HPO4). Store liquid TB at room temperature, add antibiotics just prior to use. 

LB medium Dissolve 10 g Bacto Tryptone, 5 g ofyeast extract, and 10 g of NaCl in water and adjust the volume to 1 L. Autoclave in narrow mouth flask or bottle and store at room temperature. 

Agarose top Dissolve 10 g Bacto Tryptone, 5 g yeast extract, 5 g NaCl, 1 g MgCl2-6H20 per 1 L water and add 7 g agarose. Autoclave, dispense into aliquots of 100 mL. Store as solid at room temperature, melt the medium as needed. 

Solid agar plates 0.5% yeast extract, 1% Bacto-tryptone, 0.5% NaCl,... 

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