Difference between revisions of "Part:BBa K4245209:Design"
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A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified. (Nilsson et al., 1994) | A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified. (Nilsson et al., 1994) | ||
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| − | Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (BBa_K4245006), hsa-mir-133a-3p (BBa_K4245009), and hsa-miR-451a (BBa_K4245012))— to be upregulated in correlation to CAD (Kaur et al., 2020). For | + | Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (BBa_K4245006), hsa-mir-133a-3p (BBa_K4245009), and hsa-miR-451a (BBa_K4245012))— to be upregulated in correlation to CAD (Kaur et al., 2020). For hsa-miR-451a, we designed two complementary arms, BBa_K4245106, the 3' arm for hsa-miR-451a and BBa_K4245113, the 5' arm for hsa-miR-451a. For the reporter, we decided on both BBa_K4245130/BBa_K4245132 the FAM and BHQ1 labeled linear probes, and BBa_K4245134/ BBa_K4245135, the DNA fluorescent aptamer split lettuce, due to their frugality and similar wavelength to GFP. |
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Revision as of 17:36, 11 October 2022
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Design Notes
A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified. (Nilsson et al., 1994)
Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (BBa_K4245006), hsa-mir-133a-3p (BBa_K4245009), and hsa-miR-451a (BBa_K4245012))— to be upregulated in correlation to CAD (Kaur et al., 2020). For hsa-miR-451a, we designed two complementary arms, BBa_K4245106, the 3' arm for hsa-miR-451a and BBa_K4245113, the 5' arm for hsa-miR-451a. For the reporter, we decided on both BBa_K4245130/BBa_K4245132 the FAM and BHQ1 labeled linear probes, and BBa_K4245134/ BBa_K4245135, the DNA fluorescent aptamer split lettuce, due to their frugality and similar wavelength to GFP.
Source
References
Kaur, A., Mackin, S. T., Schlosser, K., Wong, F. L., Elharram, M., Delles, C., Stewart, D. J., Dayan, N., Landry, T., & Pilote, L. (2019). Systematic review of microrna biomarkers in acute coronary syndrome and stable coronary artery disease. Cardiovascular Research, 116(6), 1113–1124. https://doi.org/10.1093/cvr/cvz302 Nilsson, M., Malmgren, H., Samiotaki, M., Kwiatkowski, M., Chowdhary, B. P., & Landegren, U. (1994). Padlock probes: Circularizing oligonucleotides for localized DNA detection. Science, 265(5181), 2085–2088. https://doi.org/10.1126/science.7522346