IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/18

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Misc.

 * Streaked TSA plates of 8325-4 and RN4220, previous ones did not have enough individual colonies to be useful
 * Autoclaved pipette tips
 * Prepared 0.1% + 0.01% Crystal Violet (by accident) - was of no use

Biofilm Growth Protocol Day 5+ - 100811a + 100811b

 * Resolubilized 100811a + b with 95% ehtanol
 * Corresponding plates were labelled 100811a-b and 100811b-b
 * Placed in biosafety cabinet, waiting for plate reader to be free
 * Took readings

Biofilm Growth Protocol Day 3 - 100816M + 100816E

 * Followed day 3 protocol

Biofilm Growth Protocol Day 4 - 100814M + 100814E

 * Stained with (what we later realized to be) 0.01% Crystal Violet
 * Took readings, did NOT resolubilize today

Readings

 * Took readings on 100811a-b, 100811b-b, 100814M and 100814E
 * All readings were garbage will place below soon
 * Decided to ethanol lid and bottom of plates before re-doing measurements

100811a-b Readings

 * Red = control

100811b-b Readings

 * Red = control

100814M Readings

 * Red = control

100814E Readings

 * Red = control

Mini-prep

 * 3T1,3T2,4T1,4T2
 * Protocol: Rafael's alkaline lysis

Nanodrop [ng/uL]:
 * 3T1: 2541.7
 * 3T2: 3053.8
 * 4T1: 3357.5
 * 4T2: 3587.8


 * Primers: VF2 and VR
 * 5uM each; 6uL in total (require 1uL of 5uM primers)

QS Track

 * Miniprepped agrCA (following Raf's alkaline lysis protocol) from distribution (after following cPCR program using UBC iGEM protocols)
 * No colonies for agrCA (natural form) showed up yesterday (both agrCA by itself and agrCA with terminator)
 * Therefore, PCR'd more agrCA.
 * Used same program, amount of reagent, and protocol as August 11 agr CA PCR.
 * Ran on gel with 1.25% agarose, 0.5X TBE, 40 minutes, 100 V
 * Bands showed up where expected (~2.1 kb)
 * Used Qiagen PCR purification kit to purify agr CA product.
 * Checked DNA quality with nanodrop. A260/280=1.90. A260/230=1.70. [DNA]=68.7 ng/μL.


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