IGEM:MIT/2007/Notebook/2007-7-7: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
 
(6 intermediate revisions by one other user not shown)
Line 2: Line 2:
#Pick plates and make liquid cultures
#Pick plates and make liquid cultures


==Checked plates==
==Checked plates 9 AM==
At 9 AM
Smears eeeeverywhere.  A lot of growth on last night's plates.


<pre>
<pre>
Line 71: Line 71:
</pre>
</pre>


==Inoculating==
==Inoculating 9AM==
Picked two colonies from and inoculated in liquid culture the following:
Picked two colonies from and inoculated in liquid culture the following:
*7/5/07 plated B0034, F2620, and E1010
*7/5/07 plated B0034, F2620, and E1010
*7/6/07 plated E1010
*7/6/07 plated E1010


Left in 37C room to incubate overnight
11:30 AM, put cultures in 37C room to incubate overnight
 
Tubes have white tape tags that read:
Tubes have white tape tags that read:
<pre>
<pre>
iGEM  7/7/07  LB+[antibiotic]
iGEM  7/7/07  LB+[antibiotic]
[Part #] in strain DH5a
[Part #] in strain DH5a
from [source plate date] [source plate #]  [pick#]
from plate [source plate date] [source plate #]  [pick#]
</pre>
 
ex:
<pre>
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
</pre>
 
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.
 
<pre>
F2620 from 7/6/07
Plate  # Colonies    Notes
1      53
2      31
3      ~60            Smear not counted
</pre>
 
 
<pre>
B0034 from 7/6/07
Plate  # Colonies    Notes
1      ~40            Smears not counted
2      120            Satellites not counted
3      ~60            Smears not counted
</pre>
</pre>
==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.)

Latest revision as of 14:43, 7 July 2007

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.)