Difference between revisions of "Team:UChicago/Notebook"

-Made a list of things to order for the future: IMPORTANT; CANNOT DO YED-ARG PLATES WITHOUT MATERIALS
-Printed most of the protocols; they are located on the lab bench when needed.
-Made LB, LB amp, and YED plates
Note: LB amp plates did not turn out as well as the others; will reattempt on 6/13/2017
-LABELLING ON PLATES
One red stripe → LB plate
One red stripe with one blue stripe → LB Amp plate
One black stripe → YED plates
-The plates have been left on the bench overnight; they will be put in the fridge tomorrow morning. Jason had cleared out a spot for us in the Glick fridge for our use!

-Task List
-Make LB and YED broth
-Program PCR protocols
-Autoclave dirty glassware
-Use pOW1 to test plating concentrations (do we have pOW1? If not, order or ask Jason?)
-Still haven’t met the glassware cleaning lady
-Mark all glassware iGem
-Redid the LB-amp plates
-Made the Drop-Out plates (-Arg)
-Talked to Shannon about cleaning our glassware
-Labelled our glassware
-The petri dishes are in the fridge (all made within the last two days)
-Messaged Steve about the pOW1 plasmid
Plan on doing pOW1 tomorrow

-Task List
-Plating Concentrations of pOW1
-The idea of plating concentrations:
Aseptic Laboratory Techniques: Plating Methods
-Learned how to use the Nanodrop
-Made two stocks of 500 mL LB media (1000 mL total)
-Spread six different DNA concentrations of the 126 ng/μl plasmid sample (AJ43) Jason gave us onto LB + amp plates
-Left these plates overnight in a 37 degrees Celsius incubator
Plate 1 = negative control (no DNA, contains 50 μl EB and 50 μl competent cells)
Plate 2 = 453.6 ng DNA (3.6 μL DNA, 46.4 μL EB, 50 μL CC)
Plate 3 = 315 ng DNA (2.5 μL DNA, 47.5 μL EB, 50 μL CC)
Plate 4 = 239.4 ng DNA (1.9 μL DNA, 48.1 μL EB, 50 μL CC)
Plate 5 = 201.6 ng DNA (1.6 μL DNA, 48.4 μL EB, 50 μL CC)
Plate 6 = 163.8 ng DNA (1.3 μL DNA, 48.7 μL EB, 50 μL CC)
-Took a small sample of pOW1 (in frozen bacteria) from stock in the -80℃ fridge
-Put the pOW1 into 5 mL of LB + amp liquid media (1 μL ampicillin / 1 mL LB)
-Left the sample in the 37 degrees shaker overnight to incubate

-Task List
-Mini-prep the pOW1
-Transform the isolated pOW1 DNA into competent cells
-Spread the competent cells on LB + amp plates
-Leave these plates overnight in a 37 degrees Celsius incubator
-Retrieved the AJ43 plates

-Mini-prepped the pOW1 that had been incubating in the shaker overnight.
The best sample we got was 61.6 ng/μL DNA
-Transformed the DNA into competent cells
Negative control = 50 μL CC
1 = 1 μL pOW1 sample, 50 μL CC
2 = 5 μL pOW1 sample, 50 μL CC
-Incubated the transformations for 2 hours in a 37℃ shaker
-Autoclaved pipette tips

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@@ Line 150: / Line 150: @@
-<div style="display: flex; ">
+<div style="display: flex; margin-top: 20px;">
   <figure><img src="https://static.igem.org/mediawiki/2017/9/97/T--UChicago--MTT.jpg" alt="Plate" width="250" height="250" style="margin-right:100px;" >
 <figcaption>Plate 1 = no colonies</figcaption>
@@ Line 162: / Line 162: @@
 </div>
-<div style="display: flex; margin-top: 40px; ">
+<div style="display: flex; margin-top: 40px;margin-bottom: 35px; ">
   <figure><img src="https://static.igem.org/mediawiki/2017/9/97/T--UChicago--MTT.jpg" alt="Plate" width="250" height="250" style="margin-right:100px;" >
 <figcaption>Plate 4 = more than 200 colonies</figcaption>
@@ Line 173: / Line 173: @@
 </figure>
 </div>
+-Mini-prepped the pOW1 that had been incubating in the shaker overnight.<br>
+&nbsp;  &nbsp;  &nbsp;  &nbsp;The best sample we got was 61.6 ng/μL DNA<br>
+-Transformed the DNA into competent cells<br>
+&nbsp;  &nbsp;  &nbsp;  &nbsp;Negative control = 50 μL CC<br>
+&nbsp;  &nbsp;  &nbsp;  &nbsp;1 = 1 μL pOW1 sample, 50 μL CC<br>
+&nbsp;  &nbsp;  &nbsp;  &nbsp;2 = 5 μL pOW1 sample, 50 μL CC<br>
+-Incubated the transformations for 2 hours in a 37&#8451; shaker<br>
+-Autoclaved pipette tips