Difference between revisions of "Part:BBa K2973017"

 
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IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its forward primer (BBa_K2973016).
 
IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its forward primer (BBa_K2973016).
  
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===Usage and Biology===
 
===Usage and Biology===
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In our project, this primer (BBa_K2973017) was used along with its forward primer (BBa_K2973016) for the amplification reactions of the IS6110 gene ([[Part:BBa_K2973009]]) that was used as a biomarker, in order to perform isothermal amplification reactions with Recombinase Polymerase Amplification (RPA). Using this method, we were able to detect quantities as low as 10 to the minus 9 nanograms in reaction times ranging from five to 20 minutes. These primers include 5' overhangs ( a T7 promoter and a trigger sequence) to allow for transcription by a T7 RNA Polymerase and translation by a toehold-switch regulated system encoding for a reporter gene (BBa_K2973007). The expected amplified product length is 136 bp.
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RPA reaction after clean up of the amplified product:
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PCR reaction:
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Revision as of 17:02, 20 October 2019


IS6110 Reverse Primer with 5' Overhang

IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its forward primer (BBa_K2973016).


Usage and Biology

In our project, this primer (BBa_K2973017) was used along with its forward primer (BBa_K2973016) for the amplification reactions of the IS6110 gene (Part:BBa_K2973009) that was used as a biomarker, in order to perform isothermal amplification reactions with Recombinase Polymerase Amplification (RPA). Using this method, we were able to detect quantities as low as 10 to the minus 9 nanograms in reaction times ranging from five to 20 minutes. These primers include 5' overhangs ( a T7 promoter and a trigger sequence) to allow for transcription by a T7 RNA Polymerase and translation by a toehold-switch regulated system encoding for a reporter gene (BBa_K2973007). The expected amplified product length is 136 bp.


RPA reaction after clean up of the amplified product:

HTML img Tag


PCR reaction:

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Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]