IGEM:Brown/2008/Notebook/Team Resistance/2008/06/17

{| width="800"
 * style="background-color: #000000"|[[Image:Team_resistance_log.png]]
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Daily Information
Created different concentrations of cells:
 * insert what you did today here

1) Put 10mL culture (pvj4) into 6, 15mL centrifuge tubes

Results: At 600nm

original culture (20 cultures)

Important Notes

 * insert what was important here

Used Beckman Centrifuge

To balance tubes:


 * Weigh the tubes that will be in one rotor (standing them up in a styrofoam block is easiest)
 * Weigh tubes that will be placed in other rotor
 * If unbalanced, put empty tube in the lighter styrofoam block and add water to tube until both block are the same mass.


 * For bacterial cells, centrifuge for about 10 minutes at a speed a little higher than 1000rpm.

To create bacteria in mid-logarithmic phase
 * Bacteria double every 20 minutes
 * An overnight culture (15hrs) should reach stationary phase (od about 10)
 * If cell culture density is known (after an overnight culture)-- dilute down to .05 .1 cell density (to make it simple-- can calculate amount of cells after a certain amount of time
 * Cells in stationary phase don't grow much -- culture od would probably be out of spec's od range (0.2-0.7) so dilute it down to the linear range and then to 0.05 or 0.1 and let incubate for about an hour

Mid-logarathmic phase: optical density less than 1-- dilute culture in stationary phase (greater than 2 or 3) to 0.05od or 0.1od and then incubate for 45 minutes to an hour  to reach mid-logarithminc phase (remember doubling time for E.Coli is once every 20 minutes)

To grow cultures, use a culture solution that is 1/4 the volume of the flask

Setbacks

 * Insert any setbacks we had today or lessons to learn from!


 * }