IGEM:IMPERIAL/2006/LabCalendar/2006-7-10

Culturing Bacteria

 * Combine together:
 * 10 mL growth medium
 * 200 uL DH5a E. coli gram negative bacteria
 * Allow to grow for a few hours until the optical density of the bacterial solution is between 0.8 and 1.0 as measured by the spectrophotometer
 * Cool the culture for a few minutes and place in an ice bath
 * Centrifuge the bacteria to obtain a bacterial pellet
 * Remove media and place on ice
 * Add 1 mL of 100 mM calcium chloride solution
 * Vortex and place back on ice bath
 * Transfer 1 mL into a microcentrifuge tube and place back on ice
 * Remove supernatant and resuspend in 100 uL of calcium chloride

Innoculation

 * Innoculate a colony of bacteria into 2 mL of ampicillin resistant growth medium

Transfering the part from the registry into the bacteria

 * We use part I13273 from well 4H (YFP Producer controleld by 3OC6HSL Receiver Device (LuxR based receiver controls production of YFP)
 * Pipette 15 uL of ultrapure water into the well and dilute
 * Remove and transfer into a microvial
 * Remove 5 uL of DNA solution and place into solution containing the bacteria
 * Leave solution for about 40 minutes
 * Heat shock the soution at 42C for 90 sec and place back on ice bath
 * Transfer the bacteria with the part onto an agar plate and spread out using a glass spreader
 * Incubate at 37C overnight