IGEM:Harvard/2007/Laboratory Notebooks/Quorum Sensing/Week 5: Difference between revisions

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(New page: ===7/16/07=== ====Colony PCR of R-P constructs==== Stephanie Colony PCR'ed the following: *[http://parts.mit.edu/registry/index.php/Part:BBa_R0052 R0052]<[http://parts.mit.edu/registry/ind...)
 
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====Dephosphorylating the J04450 and J-T====
====Dephosphorylating the J04450 and J-T====
George dephosphorylated the J04450 and the J-T with Antarctic Phosphotase and then heat inactivated the AP. Silly George, AP is for vectors not inserts. Redo it tomorrow.
[[User:GeorgeXu#Dephosphorylation_of_J04450_and_JT|George's notebook entry]]
====Liquid Cultures of R-P constructs and J-T====
George grew liquid cultures of the following:
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0011 R0011] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0340 P0340] colony #2
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0051 R0051] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0140 P0140] colony #1
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0051 R0051] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0340 P0340] colony #1
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0052 R0052] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0140 P0140] colony #2 Lower Right
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0052 R0052] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0340 P0340] colony #1 Upper Right
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0052 R0052] <[http://parts.mit.edu/registry/index.php/Part:BBa_P0340 P0340] colony #1 Lower Right
*[http://parts.mit.edu/registry/index.php/Part:BBa_J23039 J23039] <[http://parts.mit.edu/registry/index.php/Part:BBa_T9002 T9002]
The Upper/Lower Right is due to the fact that the colony PCR plate had duplicates of those colonies and it was unclear which colony was correct (at least, the PCR and gel was correct). So, George took both samples and labeled one as coming from the upper right of the plate (when viewed with the handwriting going the correct way) versus lower right.
[[User:GeorgeXu#Liquid_Cultures|George's notebook entry]]

Revision as of 12:17, 17 July 2007

7/16/07

Colony PCR of R-P constructs

Stephanie Colony PCR'ed the following:

Stephanie's notebook entry

Plate Reader (Light Scattering)

Stephanie set up another plate reader for fluorescence. One of the things she is testing is choosing ridiculous wavelengths to see if we can get light scattering data that correlates to cell growth.

Stephanie's notebook entry Results posted by Stephanie

Miniprep of the J04450 and JT constructs

In preparation for building the constitutive RFP + JT construct, George miniprepped the following parts which were grown overnight:

George's notebook entry

Digesting the J04450 and J-T

George digested the J04450 with SpeI and PstI; he also digested the J23039>T9002 with XbaI and PstI.

George's notebook entry

Sequencing the Promoters: R0011, R0051, R0052

George and Stephanie sent the R0011, R0051, and the R0052 for sequencing.

Dephosphorylating the J04450 and J-T

George dephosphorylated the J04450 and the J-T with Antarctic Phosphotase and then heat inactivated the AP. Silly George, AP is for vectors not inserts. Redo it tomorrow.

George's notebook entry

Liquid Cultures of R-P constructs and J-T

George grew liquid cultures of the following:

The Upper/Lower Right is due to the fact that the colony PCR plate had duplicates of those colonies and it was unclear which colony was correct (at least, the PCR and gel was correct). So, George took both samples and labeled one as coming from the upper right of the plate (when viewed with the handwriting going the correct way) versus lower right.

George's notebook entry

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