IGEM:MIT/2007/Notebook/2007-7-7

Agenda

1. Pick plates and make liquid cultures

Checked plates 9 AM

Smears eeeeverywhere. A lot of growth on last night's plates.

B0034 (RBS) plated on 7/5/07

Plate    # Colonies     Notes
1        19             Overgrown, satellite colonies
2        27             Overgrown, satellite colonies are starting to get too big
3        10             Satellite colonies, didn't count ones I wasn't sure of

E1010 (eRFP) plated on 7/5/07

Plate    # Colonies     Notes
1        8              Counted the ones big enough to be picked.  A lot of little speckles?
2        5              Counted the ones big enough to be picked
3        3              Counted the ones big enough to be picked

F2620 (promoter) plated on 7/5/07

Plate    # Colonies     Notes
1        13             Counted distinct ones
2        5
3        25             Satellite colonies

Positive Controls plated on 7/6/07

Plate                 # Colonies        Notes
PUC18 in LB+Amp       21                Colonies are pretty small, same as yesterday morning's.  Really small satellite colonies
PUC18 in LB           Totally covered

B0034 (RBS) plated 7/6/07

Specks, need to incubate longer before inoculating

Plate   # Colonies      Notes
1       ~100            Smear in the middle
2       ~90             These were the ones I could see, I think there are a bunch growing on the sides
3       ~140            Two smears

F2620 (promoter) plated 7/6/07

Specks, need to incubate longer before inoculating

Plate   # Colonies      Notes
1       32
2       29
3       65

E1010 (eRFP) plated 7/6/07

Plate   # Colonies      Notes
1       27              There's an almost smear. Didn't count tiny specks
2       23              Didn't count tiny specks
3       37              "                      "

Inoculating 9AM

Picked two colonies from and inoculated in liquid culture the following:

• 7/5/07 plated B0034, F2620, and E1010
• 7/6/07 plated E1010

11:30 AM, put cultures in 37C room to incubate overnight

Tubes have white tape tags that read:

iGEM  7/7/07  LB+[antibiotic]
[Part #] in strain DH5a
from plate [source plate date] [source plate #]   [pick#]

ex:

iGEM 7/7/07 LB+Kan
E1010 in strain DH5a
from plate 7/5/07 #2   pick 2

This label means:
The test tube contains LB + Kanamycin
The test tube contains bacteria with biobrick part BBa_E1010
The source plate is labeled with the date 7/5/07 #2 and the corresponding biobrick part
The test tube contains the second colony picked from the source plate

Sorry if this is confusing guys, our source plates don't have a naming system yet.

Checked Plates 5 PM

Colonies look bigger

Smears are probably from leaving a puddle after plating.

F2620 from 7/6/07
Plate   # Colonies     Notes
1       53
2       31
3       ~60            Smear not counted

B0034 from 7/6/07
Plate   # Colonies     Notes
1       ~40            Smears not counted
2       120            Satellites not counted
3       ~60            Smears not counted

Making liquid cultures 5 PM

• Made liquid cultures in big test tubes of two colonies from each plate (7.6 #1, #2 and #3 of F2620 and B0034 -- both Amp) -- 12 tubes total
• Put in 37° room on rotater for overnight incubation
• Random Note: iGEM team is authorized to pick up the lab phone! (Just in case...Jess and I didn't pick up because we didn't know if we were allowed to today, and Francois was actually locked out of lab for a bit...oops. Sorry.)