Difference between revisions of "Part:BBa K4784002:Experience"

(→‎Applications of BBa_K4784002)
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     Add 30μl of culture fluid to 3 ml of LB-medium containing ampicillin and 0-1.5μmol of IPTG. <br>　 Cultured for 4-5h at 37℃ in culture tubes in a shaking water bath at 168 min^-1  
 
     Add 30μl of culture fluid to 3 ml of LB-medium containing ampicillin and 0-1.5μmol of IPTG. <br>　 Cultured for 4-5h at 37℃ in culture tubes in a shaking water bath at 168 min^-1  
 
5. Measure the cocentration of Pyruvate every hour
 
5. Measure the cocentration of Pyruvate every hour
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     * According to the protocol offered by assay kit's manifacture, make the standard curve. <br>    Since LB medium might contains acetate and absorbrance, acetate samples used in calibration was made by dliting acetate <br>    Standard with LB mediums instead of water.
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     The protocols is here; https://docs.aatbio.com/products/protocol-and-product-information-sheet-pis/protocol-for-amplite-colorimetric-pyruvate-assay-kit-version-5766863da6.pdf
 
     The protocols is here; https://docs.aatbio.com/products/protocol-and-product-information-sheet-pis/protocol-for-amplite-colorimetric-pyruvate-assay-kit-version-5766863da6.pdf
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    *  Sampleing  culture medium and mix with the working reagent of assay kit.
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===User Reviews===
 
===User Reviews===
 
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Revision as of 15:22, 12 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_K4784002

Experiment
1. Gibson assembly 

   Forming the plasmid contains Pdr12 and AlaDH.

2. Transformation 

   Following the Single Tube Transformation Protocol provided by iGEM, except for the process of heat shock. 
   We conducted heat-shock for 30sec at 42°C on a heat block.

3. Culturing 

   Pick colonies and culture them in LB-cam containing ampicillin overnight.

4. IPTG induction 

   Add 30μl of culture fluid to 3 ml of LB-medium containing ampicillin and 0-1.5μmol of IPTG. 
　 Cultured for 4-5h at 37℃ in culture tubes in a shaking water bath at 168 min^-1

5. Measure the cocentration of Pyruvate every hour 

   The protocols is here; https://docs.aatbio.com/products/protocol-and-product-information-sheet-pis/protocol-for-amplite-colorimetric-pyruvate-assay-kit-version-5766863da6.pdf

User Reviews

UNIQ48e5e92a09a6360b-partinfo-00000000-QINU UNIQ48e5e92a09a6360b-partinfo-00000001-QINU