Difference between revisions of "Part:BBa K103001:Experience"

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===Applications of BBa_K103001===
 
===Applications of BBa_K103001===
*Introduction of mutations to specific region of DNA.  
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===Increasing intracellular mutation level===
 
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<partinfo>BBa_K103001 AddReview 5</partinfo>
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<I>Keton</I>
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We have introduced aid gene to E. coli and measured level of mutations using rifampicin test. It works. The results are here:
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*[http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=19_May_2008&arg1=20_May_2008&arg2=21_May_2008&name=Rifampicin%20test%20%231 Rifampicin test #1]
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*[http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=22_May_2008&arg1=23_May_2008&arg2=24_May_2008&name=Rifampicin%20test%20%232 Rifampicin test #2]
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*[http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=28_May_2008&arg1=29_May_2008&arg2=2_June_2008&name=Rifampicin%20test%20%233 Rifampicin test #3]
  
===User Reviews===
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===Introduction of mutations to specific region of DNA===
 
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<I>Keton</I>
 
<I>Keton</I>
 
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We have tested this part in conjunction with T7 polymerase and target gene (lacZ) under T7 promoter in Top10 strain. Although we have obtained white colonies in various experimental setups sequencing revealed no mutations in target region (T7 promoter or lacZ ORF). We have repeated this experiment in GM2163 strain which is incapable of MutHLS mediated DNA repair due to lack of Dam and Dcm methyltransferases and obtained heritable white phenotype at high frequencies. Unfortunately we have no sequencing data to prove that mutations really occurred.
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We have tested this part in conjunction with T7 polymerase and target gene (lacZ) under T7 promoter in Top10 strain. Although we have obtained white colonies in various experimental setups ([http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=2_June_2008&arg1=5_June_2008&arg2=6_June_2008&arg3=11_June_2008&arg4=12_June_2008&arg5=13_June_2008&arg6=16_June_2008&name=Blue/white%20and%20rifampicin%20test%20%231 #1] and [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=4&arg0=17_June_2008&arg1=18_June_2008&arg2=19_June_2008&arg3=23_June_2008&name=Blue/white%20and%20rifampicin%20test%20%232 #2]) sequencing revealed no mutations in target region (T7 promoter or lacZ ORF). We have [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=4&arg0=29_August_2008&arg1=30_August_2008&arg2=31_August_2008&arg3=1_September_2008&name=Blue-white%20screening%20and%20rifampicin%20test%20in%20GM2163 repeated] this experiment in GM2163 strain which is incapable of MutHLS mediated DNA repair due to lack of Dam and Dcm methyltransferases and obtained heritable white phenotype at high frequencies. Unfortunately we have no sequencing data to prove that mutations really occurred.
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Revision as of 19:47, 27 October 2008

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_K103001

Increasing intracellular mutation level

•••••

Keton

We have introduced aid gene to E. coli and measured level of mutations using rifampicin test. It works. The results are here:

  • [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=19_May_2008&arg1=20_May_2008&arg2=21_May_2008&name=Rifampicin%20test%20%231 Rifampicin test #1]
  • [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=22_May_2008&arg1=23_May_2008&arg2=24_May_2008&name=Rifampicin%20test%20%232 Rifampicin test #2]
  • [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=3&arg0=28_May_2008&arg1=29_May_2008&arg2=2_June_2008&name=Rifampicin%20test%20%233 Rifampicin test #3]

Introduction of mutations to specific region of DNA

UNIQd146430d8e592e16-partinfo-00000001-QINU

•••••

Keton

We have tested this part in conjunction with T7 polymerase and target gene (lacZ) under T7 promoter in Top10 strain. Although we have obtained white colonies in various experimental setups ([http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=2_June_2008&arg1=5_June_2008&arg2=6_June_2008&arg3=11_June_2008&arg4=12_June_2008&arg5=13_June_2008&arg6=16_June_2008&name=Blue/white%20and%20rifampicin%20test%20%231 #1] and [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=4&arg0=17_June_2008&arg1=18_June_2008&arg2=19_June_2008&arg3=23_June_2008&name=Blue/white%20and%20rifampicin%20test%20%232 #2]) sequencing revealed no mutations in target region (T7 promoter or lacZ ORF). We have [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=4&arg0=29_August_2008&arg1=30_August_2008&arg2=31_August_2008&arg3=1_September_2008&name=Blue-white%20screening%20and%20rifampicin%20test%20in%20GM2163 repeated] this experiment in GM2163 strain which is incapable of MutHLS mediated DNA repair due to lack of Dam and Dcm methyltransferases and obtained heritable white phenotype at high frequencies. Unfortunately we have no sequencing data to prove that mutations really occurred.


UNIQd146430d8e592e16-partinfo-00000003-QINU