Difference between revisions of "Part:BBa K4664004"

 
 
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<partinfo>BBa_K4664004 parameters</partinfo>
 
<partinfo>BBa_K4664004 parameters</partinfo>
 
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===Background===
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Our reporter is composed of a fluorophore linked to a quencher molecule. While the two are connected, the quencher absorbs energy from the fluorophore, thus preventing it from emitting fluorescence. But when cas12a trans-cleavage acts on the quencher-fluorophore and severs their link, the fluorophore would start giving out fluorescence. Thus, the fluorescence level is used to quantify the concentration of miRNA biomarkers in the sample tested.
 +
<br>
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===Design===
 +
Sequence:
 +
<br>5-FAM-TTATT-IABkFQ-3’
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===Results===
 +
We performed a series of standard MB-ERC2 reactions with different reporter concentrations at miRNA concentration levels 1 pM and 10 pM. Figures 1-3 show results from these trials.
 +
<br>
 +
https://static.igem.wiki/teams/4664/wiki/part/p464004-1.png
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<br>
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<i>Figure 1. Reporter 1 pM.</i>
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<br>
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https://static.igem.wiki/teams/4664/wiki/part/p464004-2.png
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<br>
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<i>Figure 2. Reporter 10 pM.</i>
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<br>
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https://static.igem.wiki/teams/4664/wiki/part/p464004-3.png
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<br>
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<i>Figure 3. Reporter final values.</i>
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<br>
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===References===
 +
Jin, J., Vaud, S., Zhelkovsky, A., Pósfai, J., & McReynolds, L. A. (2016). Sensitive and specific miRNA detection method using SplintR Ligase. Nucleic Acids Research, 44(13), e116. https://doi.org/10.1093/nar/gkw399
 +
<br>
 +
He, Y., Wen, Y., Tian, Z., Hart, N. T., Han, S., Hughes, S. J., & Zeng, Y. (2023b). A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs. Nature Biomedical Engineering. https://doi.org/10.1038/s41551-023-01033-1

Latest revision as of 06:18, 12 October 2023


Reporter

The reporter is used for showing the fluorescence signal.

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]


Background

Our reporter is composed of a fluorophore linked to a quencher molecule. While the two are connected, the quencher absorbs energy from the fluorophore, thus preventing it from emitting fluorescence. But when cas12a trans-cleavage acts on the quencher-fluorophore and severs their link, the fluorophore would start giving out fluorescence. Thus, the fluorescence level is used to quantify the concentration of miRNA biomarkers in the sample tested.

Design

Sequence:
5-FAM-TTATT-IABkFQ-3’

Results

We performed a series of standard MB-ERC2 reactions with different reporter concentrations at miRNA concentration levels 1 pM and 10 pM. Figures 1-3 show results from these trials.
p464004-1.png
Figure 1. Reporter 1 pM.
p464004-2.png
Figure 2. Reporter 10 pM.
p464004-3.png
Figure 3. Reporter final values.

References

Jin, J., Vaud, S., Zhelkovsky, A., Pósfai, J., & McReynolds, L. A. (2016). Sensitive and specific miRNA detection method using SplintR Ligase. Nucleic Acids Research, 44(13), e116. https://doi.org/10.1093/nar/gkw399
He, Y., Wen, Y., Tian, Z., Hart, N. T., Han, S., Hughes, S. J., & Zeng, Y. (2023b). A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs. Nature Biomedical Engineering. https://doi.org/10.1038/s41551-023-01033-1