Difference between revisions of "Part:BBa J01010:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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<I>HZAU-China 2014</I>
 
<I>HZAU-China 2014</I>
 
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We want to know under the participation of taRNA and crRNA, whether can reduce leakage of lac promoter’s expression. So we did some experiment to text. First, built the tow devices and from top to bottom Numbers for 1 2 , and then, respectively transform the three plasmid into BL21 competent cell. Pick monoclonal colony. Add 5ml LB liquid and incubate at 37 Celsius degree for 8 hours. Secondly, add 5ul of E.coli liquid cultural medium from each tube to two new tubes named A, B respectively with 5ml M9 broth in it. Add 0.1ul IPTG to all A tubes (1A, 2A, ) while add nothing to all B tubes (1B, 2B, ). Incubating at 37 Celsius degree for 4 hours. Lastly, easuring the fluorescent degree by utilizing the enzyme-labeled instrument.
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We want to know under the participation of taRNA and crRNA, whether can reduce leakage of lac promoter’s expression. So we did some experiment to text. First, built the tow devices and from top to bottom Numbers for 1 2 , and then, respectively transform the two plasmid into BL21 competent cell. Pick single colonies. Add 5ml LB liquid and incubate at 37 Celsius degree for 8 hours. Secondly, add 5ul of E.coli liquid cultural medium from each tube to two new tubes named A, B respectively with 5ml M9 broth in it. Add 0.1ul IPTG to all A tubes (1A, 2A, ) while add nothing to all B tubes (1B, 2B, ). Incubating at 37 Celsius degree for 4 hours. Lastly, easuring the fluorescent degree by utilizing the enzyme-labeled instrument.
 
Conclusion : riboregulators ensure much lower leakage of the promoters.  
 
Conclusion : riboregulators ensure much lower leakage of the promoters.  
  
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[[File:HZAU2014-图片1.jpg|700px|right]]
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[[File:HZAU2014-图片1.jpg|380px|left]]
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[[File:HZAU2014-ribo.jpg|700px]]
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[[File:HZAU2014-ribo.jpg|800px]]
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<partinfo>BBa_J01008 AddReview 3</partinfo>
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<I>Hong_Kong_HKUST_2014</I>
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We saw a strong repression after adding cis-repressing sequence 5’ of the RBS. After the induction of arabinose to activate the Pbad promoter that regulates the expression of the key. we saw around 13-fold increase. This increase however, only corresponded to 0.4% of the fluorescence of samples that were unrepressed.
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For detailed information and figures, visit HKUST iGEM 2014 team wiki page[[http://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization]].
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Latest revision as of 09:30, 18 October 2014

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Please enter how you used this part and how it worked out.

Applications of BBa_J01010

User Reviews

UNIQ46468fd6db072512-partinfo-00000000-QINU UNIQ46468fd6db072512-partinfo-00000001-QINU

•••••

HZAU-China 2014

We want to know under the participation of taRNA and crRNA, whether can reduce leakage of lac promoter’s expression. So we did some experiment to text. First, built the tow devices and from top to bottom Numbers for 1 2 , and then, respectively transform the two plasmid into BL21 competent cell. Pick single colonies. Add 5ml LB liquid and incubate at 37 Celsius degree for 8 hours. Secondly, add 5ul of E.coli liquid cultural medium from each tube to two new tubes named A, B respectively with 5ml M9 broth in it. Add 0.1ul IPTG to all A tubes (1A, 2A, ) while add nothing to all B tubes (1B, 2B, ). Incubating at 37 Celsius degree for 4 hours. Lastly, easuring the fluorescent degree by utilizing the enzyme-labeled instrument. Conclusion : riboregulators ensure much lower leakage of the promoters.

HZAU2014-图片1.jpg
HZAU2014-图片2.jpg

HZAU2014-ribo.jpg

•••

Hong_Kong_HKUST_2014


We saw a strong repression after adding cis-repressing sequence 5’ of the RBS. After the induction of arabinose to activate the Pbad promoter that regulates the expression of the key. we saw around 13-fold increase. This increase however, only corresponded to 0.4% of the fluorescence of samples that were unrepressed.

For detailed information and figures, visit HKUST iGEM 2014 team wiki pagehttp://2014.igem.org/Team:Hong_Kong_HKUST/riboregulator/characterization.