ออก. Insert the two-way connector attached to a length of 3/16" inner-diameter aquarium tubing. Open the main gas tank valve and set the regulator to 2 lb with the regulator adjusting screw. Place a sterile 1 ml plastic pipet in other end of the aquarium tubing, and keep pipet tip sterile by placing the tip end in a sterile bag (figures 1 & 2). Use a new pipet for each bag. Heat seal each filled bag (metalized poly pouch) with a bag sealer. Cut a very small corner from the top of the sealed end. Squeeze air from bag by pressing area above liquid until the area is flat. Insert pipet into open corner of bag and open on-off valve on the gas hose. Fill area above the broth with gas. Repeat squeezing and gassing each bag 2 more times, ending with a gassing step. Quickly heat seal or tape the corner of the bag shut. Place gassed bags into baskets lined with plastic bags. Set the basket(s) onto a shaker incubator platform.Set shaker speed to 175-200 rpm.