Difference between revisions of "Team:UChicago/Notebook"

·Made a list of things to order for the future
·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℃ 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℃ 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℃ 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

-Retrieved the pOW1 plates

-pOW1 plating concentrations for next week:
1 μL pOW1 + 49 μL EB + 50 μL CC
2.5 μL pOW1 + 47.5 μL EB + 50 μL CC
5 μL pOW1 + 45 μL EB + 50 μL CC

-Primers either to arrive on Tuesday or Wednesday
-Transformed the pOW1 DNA into competent cells
1 = Negative control = 50 μL CC
2 = 1 μL pOW1 + 49 μL EB + 50 μL CC
3 = 2.5 μL pOW1 + 47.5 μL EB + 50 μL CC
4 = 5 μL pOW1 + 45 μL EB + 50 μL CC
-Incubated the transformations for 2 hours in a 37℃ shaker
-Spread the samples on LB + amp plates
-Left the plates overnight in a 37℃incubator
-Tasks for tomorrow:
-Check on the plates
-If the primers have arrived, start Workflow 1!

-Observed the plates from the previous transformation
-Retrieved pOW1 plates

-Transformed the pOW1 DNA again into competent cells
1 = Negative control = 50 μL CC
2 = 0.5 μL pOW1 + 50 μL CC
3 = 10 μL pOW1 + 40 μL EB + 50 μL CC
4 = 15 μL pOW1 + 35 μL EB + 50 μL CC
5 = 20 μL pOW1 + 30 μL EB + 50 μL CC
-Cool website that gives you quick-change primers (by just entering the sequence you want to amplify):
QuikChange Primer Design
-Plated autotrophic (arg) yeast and non-autotrophic yeast on -Arg and +Arg
Incubated the two plates in 30℃ overnight
-PCR protocol for pSB1C3mut:

-Retrieved the yeast plates:

-Retrieved pOW1 plates:

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July-----

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August-----

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@@ Line 84: / Line 84: @@
 <button class="accordion">6/12/2017</button>
 <div class="panel">
-   <p>-Made a list of things to order for the future: IMPORTANT; CANNOT DO YED-ARG PLATES WITHOUT MATERIALS<br>
+   <p>&#183;Made a list of things to order for the future<br>
--Printed most of the protocols; they are located on the lab bench when needed. <br>
+&#183;Printed most of the protocols; they are located on the lab bench when needed<br>
--Made LB, LB amp, and YED plates<br>
+&#183;Made LB, LB amp, and YED plates<br>
-  &nbsp;  &nbsp;  &nbsp;  &nbsp;Note: LB amp plates did not turn out as well as the others; will reattempt on 6/13/2017 <br>
+  &nbsp;  &nbsp;  &nbsp;  &nbsp;&#183;Note: LB amp plates did not turn out as well as the others; will reattempt on 6/13/2017 <br>
--LABELLING ON PLATES<br>
+&#183;LABELLING ON PLATES<br>
-  &nbsp;  &nbsp;  &nbsp;  &nbsp; One red stripe → LB plate <br>
+  &nbsp;  &nbsp;  &nbsp;  &nbsp; &#183;One red stripe &#8594; LB plate <br>
-&nbsp;  &nbsp;  &nbsp;  &nbsp; One red stripe with one blue stripe → LB Amp plate
+&nbsp;  &nbsp;  &nbsp;  &nbsp; &#183;One red stripe with one blue stripe &#8594; LB Amp plate
   <br>
-&nbsp;  &nbsp;  &nbsp;  &nbsp; One black stripe → YED plates
+&nbsp;  &nbsp;  &nbsp;  &nbsp; &#183;One black stripe &#8594; YED plates
   <br>
--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!
+&#183;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!
 </p>
 </div>