Team:Mississippi State/Making Competent Cells


 * To make competent cells(XL1-Blue), do exactly as follows:


 * 1) Add 2ml LB Broth to Culture Tube.
 * 2) Add 1ul antibiotic for every 1ml LB(in this case, 2ul of Tetracycline).
 * 3) Get cell stock from -80C freezer.
 * 4) Scrape cells from top with tip of pipette.
 * 5) Add to LB Broth.
 * 6) Label (Culture name, date, iGEM)
 * 7) Put in 37C shaker overnight.
 * 8) Remove next morning.
 * 9) Get pipette gun from 1st drawer from left under lab bench closest to door.
 * 10) Get 10ml tube from lab bench opposite 1st lab bench.
 * 11) Add 25ml LB Broth to autoclaved flask(KEEP THE FOIL COVER!).
 * 12) Add 25ul Tet (in IGEM 2007 Box, -20C freezer)
 * 13) Add 250ul cells from overnight culture.
 * 14) Cover with aluminum
 * 15) Put in 37C shaker for 2hr.
 * 16) Check cell density
 * 17) Pour into centrifuge tubes.
 * 18) Centrifuge 10-15min. in SS34 model, which is setting 05 for 15min at 5000rpm and +4C.
 * 19) Pour out supernatant into same flask, autoclave this flask before disposal.
 * 20) Resuspend cells in CaCl, using half previous volume: for example, if you originally had cells in 10ml LB, you would use 5ml CaCl.
 * 21) Centrifuge again at same settings.
 * 22) Pour out supernatant.
 * 23) Resuspend in CaCl, this time using 2ml of CaCl.
 * 24) Store on Ice for 1hr.