Difference between revisions of "Part:BBa K4202025:Experience"

(Characterization)
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<b>Characterization method</b>
 
<b>Characterization method</b>
<p>The transformant were inoculate to 2ml LB medium and cultured in the test tube overnight. The next day, the culture were inoculated into 2ml TB medium containing 0%, 0.1%, 0.5%, 1%, 2%, 5% sucrose (w/v) at the ratio of 1:100. After cultured in 37℃, 220rpm for 7 hours, the culture were diluted into PBS at the ratio of 1:1000 and were measured  the fluorescence intensity through flow cytometry.</p>
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<p>The transformant were inoculate to 2ml LB medium and cultured in the test tube overnight. The next day, the culture were inoculated into 2ml TB medium containing 0%, 0.1%, 0.5%, 1%, 2%, 5% sucrose (w/v) at the ratio of 1:100. After cultured in 37℃, 220rpm for 7 hours, the culture were diluted into PBS at the ratio of 1:1000 and were measured  the fluorescence intensity through flow cytometry. The cluster of <i>Bacillus subtilis</i> should be easily gated from SSC-A vs. FSH-H figure. </p>
 
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<div align="center">[[File:Gate.jpg|500px]]</div>
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<div align="center"><b>Fig 1</b> The <i>Bacillus subtilis</i> in the SSC-A vs. FSH-H figure</div>
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 14:31, 12 October 2022


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K4202025


Characterization


Vector construct

To characterization this part, we constructed the measurement device BBa_K4202033 and then constructed it to the pHY300PLK plasmid. The vector was then transformed into Bacillus subtilis WB600 strain.


Characterization method

The transformant were inoculate to 2ml LB medium and cultured in the test tube overnight. The next day, the culture were inoculated into 2ml TB medium containing 0%, 0.1%, 0.5%, 1%, 2%, 5% sucrose (w/v) at the ratio of 1:100. After cultured in 37℃, 220rpm for 7 hours, the culture were diluted into PBS at the ratio of 1:1000 and were measured the fluorescence intensity through flow cytometry. The cluster of Bacillus subtilis should be easily gated from SSC-A vs. FSH-H figure.


Gate.jpg
Fig 1 The Bacillus subtilis in the SSC-A vs. FSH-H figure

User Reviews

UNIQ25459be7fee713bb-partinfo-00000001-QINU UNIQ25459be7fee713bb-partinfo-00000002-QINU