Difference between revisions of "Part:BBa K4630114:Experience"

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===Applications of BBa_K4630114===
 
===Applications of BBa_K4630114===
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This part is the more integrated form that contains three cassettes. Using this part, we can verify the concept of multi-level recording and gain comprehensive data.
 +
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===General Induction Protocol===
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# Pick a colony from the verified plate, cultivate in 5ml liquid LB media for 12 hours.
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# Transfer 100µl bacteria solution to 3ml new liquid LB media. Cultivate to an OD600 of 0.3~0.5.
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# Add L-Arabinose and IPTG to final concentrations of 2g/L and 3g/L, and induce for 22 hours and 5 hours, respectively.
 +
# Spread the diluted solution to plates with appropriate antibiotics and cultivate for 12 hours.
 +
# Pick the colonies and test the editing result.
 +
 +
 +
===Result===
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====Proof-of-Concept===
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We implement induction over the part, and it is successfully knock-out.
 
We implement induction over the part, and it is successfully knock-out.
 
<html>
 
<html>
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Also, we performed time-gradient tests over the part.
 
Also, we performed time-gradient tests over the part.
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<html>
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        <div class="col-lg" style="margin:auto;text-align:center;">
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                <img style="margin:20px auto 5px auto;" src="https://static.igem.wiki/teams/4630/wiki/parts/https://static.igem.wiki/teams/4630/wiki/parts/bba-k4630100-stgrna-barcode-cassette-1/table10.png" width="80%">
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</div>
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<div class="col-lg" style="margin:auto;text-align:left;">
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                <p style="color:Gray; padding:0px 30px 10px;">
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Fig 2 Time-gradient test over the part<br>
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</p>
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</div>
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</html>
  
 
===User Reviews===
 
===User Reviews===

Revision as of 23:43, 11 October 2023


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_K4630114

This part is the more integrated form that contains three cassettes. Using this part, we can verify the concept of multi-level recording and gain comprehensive data.

General Induction Protocol

  1. Pick a colony from the verified plate, cultivate in 5ml liquid LB media for 12 hours.
  2. Transfer 100µl bacteria solution to 3ml new liquid LB media. Cultivate to an OD600 of 0.3~0.5.
  3. Add L-Arabinose and IPTG to final concentrations of 2g/L and 3g/L, and induce for 22 hours and 5 hours, respectively.
  4. Spread the diluted solution to plates with appropriate antibiotics and cultivate for 12 hours.
  5. Pick the colonies and test the editing result.


Result

=Proof-of-Concept

We implement induction over the part, and it is successfully knock-out.

Fig 1 The successful knock-out of the stgRNA-cassette (1+2+3)

Also, we performed time-gradient tests over the part.

Fig 2 Time-gradient test over the part

User Reviews

UNIQ53a61eab470e8817-partinfo-00000002-QINU UNIQ53a61eab470e8817-partinfo-00000003-QINU