Team:Lethbridge/Notebook/Lab Work/October

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(Difference between revisions)
(→October 16, 2010)
(→JV)
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<b>Objective:</b> Separate different fractions of the cytosol of cells containing catechol 2,3-dioxygenase <br>
<b>Objective:</b> Separate different fractions of the cytosol of cells containing catechol 2,3-dioxygenase <br>
<b>Method:</b>
<b>Method:</b>
-
*Grow DH5&alpha;<br>
+
*Grow DH5&alpha; cells in 500mL of LB media with Amp<sup>+</sup>.
 +
*Optical density of cell suspension (measured at 600nm): 4.55
 +
*Spun cells down: 10 minutes, 4<sup>o</sup>C, at 5000RPM
 +
*Cell pellet weight is 3.03g
 +
*Flash froze cells and stored at -80<sup>o</sup>C <br>
 +
 
 +
Plasmid DNA was also isolated using QIAGEN spin column protocol. DNA was eluted to 60&micro;L.<br>

Revision as of 00:03, 19 October 2010



Feel free to look around our notebook!

Here you can check out the work we have done in the lab, click on a month to take a look!

Contents

October

October 2, 2010

ADS

Objective: Purify plasmid DNA from transformed Mr. GENE synthesized DNA

  • <partinfo>K331022</partinfo>
  • <partinfo>K331023</partinfo>
  • <partinfo>K331024</partinfo>
  • <partinfo>K331025</partinfo>

Method: Use Qiagen miniprep method with BioBasic EZ-10 Spin columns.

Objective: Create 20 new biobricks using recently arrived synthesized DNA and terminator <partinfo>B0015</partinfo>, pLac <partinfo>R0010</partinfo>, and pTet <partinfo>R0040</partinfo> to create the following new biobricks:

  • 1)R0010 + K331022 to obtain <partinfo>K331026</partinfo>
  • 2)R0040 + K331022 to obtain <partinfo>K331030</partinfo>
  • 3)K331022 + B0015 to obtain <partinfo>K331034</partinfo>
  • 4)K331022 + K331024 to obtain (NAME BIOBRICK!!!!)
  • 5)R0010 + K331023 to obtain <partinfo>K331027</partinfo>
  • 6)R0040 + K331023 to obtain <partinfo>K331031</partinfo>
  • 7)K331023 + B0015 to obtain <partinfo>K331035</partinfo>
  • 8)K331023 + K331025 to obtain (NAME BIOBRICK!!!!)
  • 9)R0010 + K331024 to obtain <partinfo>K331028</partinfo>
  • 10)R0040 + K331024 to obtain <partinfo>K331032</partinfo>
  • 11)K331024 + B0015 to obtain <partinfo>K331036</partinfo>
  • 12)K331024 + K331022 to obtain (NAME BIOBRICK!!!!)
  • 13)R0010 + K331025 to obtain <partinfo>K331029</partinfo>
  • 14)R0040 + K331025 to obtain <partinfo>K331033</partinfo>
  • 15)K331025 + B0015 to obtain <partinfo>K331037</partinfo>
  • 16)K331025 + K331023 to obtain (NAME BIOBRICK!!!!)
  • 17)K331022 to obtain <partinfo>K331022</partinfo>
  • 18)K331023 to obtain <partinfo>K331023</partinfo>
  • 19)K331024 to obtain <partinfo>K331024</partinfo>
  • 20)K331025 to obtain <partinfo>K331025</partinfo>

All parts will be inserted into pSB1C3.
Method: Use BioBrick Assembly Method
Results: Obtained RESULTS!!! positive colonies.

October 5, 2010

ADS

Objective: Purify plasmid DNA from successfully assembled biobricks.
Method: Use Qiagen miniprep method with BioBasic EZ-10 columns.
Will send these minipreps for sequencing at Macrogen.




October 16, 2010

JV

Objective: Separate different fractions of the cytosol of cells containing catechol 2,3-dioxygenase
Method:

  • Grow DH5α cells in 500mL of LB media with Amp+.
  • Optical density of cell suspension (measured at 600nm): 4.55
  • Spun cells down: 10 minutes, 4oC, at 5000RPM
  • Cell pellet weight is 3.03g
  • Flash froze cells and stored at -80oC

Plasmid DNA was also isolated using QIAGEN spin column protocol. DNA was eluted to 60µL.

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