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.)