Difference between revisions of "Part:BBa K419020:Experience"

 
(Applications of BBa_K419020)
 
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===Applications of BBa_K419020===
 
===Applications of BBa_K419020===
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'''Verification of the Part BBa_K419020'''
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[[image:photo10.jpg]]
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The DNA fragment of nhaA promoter was amplified from the part BBa_K116002 by PCR method. The nhaA PCR products (274 bp) were purified and digested with EcoRI and PstI, then ligated with EcoRI-PstI cut pSB1C3, the iGEM 2010 standard backbone. We got the pSB1C3-nhaA plasmid successfully. Furthermore, the Lux I gene with ribosome binding site (RBS) DNA was amplified from the part BBa_C0161. The Lux I PCR products (585 bp) were digested with Xba I and Pst I, then ligated with the Spe I-Pst I treated pSB1C3-nhaAvector. After E. coli transformation and plasmid preparation, the part BBa_K419020 for sensing system in our Neutrient Synthesizer was verified by EcoRI and PstI enzyme digestion in 1% agarose gel electrophoresis. The arrow (red) indicates the nhaA-Lux I DNA fragment (859 bp). The sensing system composite was transformed into E. coli for further experiments.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 17:28, 6 November 2010

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_K419020

Verification of the Part BBa_K419020

Photo10.jpg The DNA fragment of nhaA promoter was amplified from the part BBa_K116002 by PCR method. The nhaA PCR products (274 bp) were purified and digested with EcoRI and PstI, then ligated with EcoRI-PstI cut pSB1C3, the iGEM 2010 standard backbone. We got the pSB1C3-nhaA plasmid successfully. Furthermore, the Lux I gene with ribosome binding site (RBS) DNA was amplified from the part BBa_C0161. The Lux I PCR products (585 bp) were digested with Xba I and Pst I, then ligated with the Spe I-Pst I treated pSB1C3-nhaAvector. After E. coli transformation and plasmid preparation, the part BBa_K419020 for sensing system in our Neutrient Synthesizer was verified by EcoRI and PstI enzyme digestion in 1% agarose gel electrophoresis. The arrow (red) indicates the nhaA-Lux I DNA fragment (859 bp). The sensing system composite was transformed into E. coli for further experiments.

User Reviews

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