Difference between revisions of "Part:BBa K3593004"
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<partinfo>BBa_K3593004 short</partinfo> | <partinfo>BBa_K3593004 short</partinfo> | ||
| − | The probe complementary to control line and tail of aptamer amplicons in our RPA-competitive-ALFA(aptamer lateral flow assay) test strip. The SH end can react with Au nano particle thus conjugate with the particle, lymphosized on conjugation pad for reporting of red color afterwards | + | The probe complementary to control line and tail of aptamer amplicons in our RPA-competitive-ALFA(aptamer lateral flow assay) test strip. The SH end can react with Au nano particle thus conjugate with the particle, lymphosized on conjugation pad for reporting of red color afterwards. |
| − | + | ||
=Sequence and features= | =Sequence and features= | ||
<partinfo>BBa_K3593004 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3593004 SequenceAndFeatures</partinfo> | ||
=Characterisation of this part= | =Characterisation of this part= | ||
==Design== | ==Design== | ||
| − | + | Besides aptamers, we have some short single stranded DNA known as 'probes', which binds to other single stranded DNA. BBa_K3593004 is one of those so-called 'probes' in our design, and play important roles in our hardware, in which it binds to the amplicons in RPA and gives colour changes that users can notice and verify whether toxin exists or not. It has a strand consisting of a lot of thymine as spacer, as well as a sequence that can bind strongly with our aptamer with a complementary strand to it. Its 3' end is conjugated to a C6 spacer and a thiol group[1], to be conjugated to Au nanoparticles. When these probes bind with aptamer immobilized on the test line by "poly T"[2][3][4], the Au nanoparticles aggregates to show a colour change from colorless to red, reporting at the test line. | |
=References= | =References= | ||
| + | 1.Jauset-Rubio M, Svobodová M, Mairal T, et al. Aptamer Lateral Flow Assays for Ultrasensitive Detection of β-Conglutin Combining Recombinase Polymerase Amplification and Tailed Primers. Anal Chem. 2016;88(21):10701-10709. doi:10.1021/acs.analchem.6b03256<br> | ||
| + | 2.Chao Zhu, Guilan Zhang, Yafei Huang, Shuming Yang, Shuyue Ren, Zhixian Gao, Ailiang Chen, Dual-competitive lateral flow aptasensor for detection of aflatoxin B1 in food and feedstuffs, Journal of Hazardous Materials https://doi.org/10.1016/j.jhazmat.2017.10.026 <br> | ||
| + | 3.Wu, S., Liu, L., Duan, N. et al. A test strip for ochratoxin A based on the use of aptamer-modified fluorescence upconversion nanoparticles. Microchim Acta 185, 497 (2018). https://doi.org/10.1007/s00604-018-3022-0<br> | ||
| + | 4.Delley CL, Liu L, Sarhan MF, Abate AR. Combined aptamer and transcriptome sequencing of single cells. Sci Rep. 2018 Feb 13;8(1):2919. doi: 10.1038/s41598-018-21153-y. PMID: 29440771; PMCID: PMC5811598.<br> | ||
| + | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Latest revision as of 03:27, 28 October 2020
modified ssDNA, Au nano particle reporter probe
The probe complementary to control line and tail of aptamer amplicons in our RPA-competitive-ALFA(aptamer lateral flow assay) test strip. The SH end can react with Au nano particle thus conjugate with the particle, lymphosized on conjugation pad for reporting of red color afterwards.
Sequence and features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterisation of this part
Design
Besides aptamers, we have some short single stranded DNA known as 'probes', which binds to other single stranded DNA. BBa_K3593004 is one of those so-called 'probes' in our design, and play important roles in our hardware, in which it binds to the amplicons in RPA and gives colour changes that users can notice and verify whether toxin exists or not. It has a strand consisting of a lot of thymine as spacer, as well as a sequence that can bind strongly with our aptamer with a complementary strand to it. Its 3' end is conjugated to a C6 spacer and a thiol group[1], to be conjugated to Au nanoparticles. When these probes bind with aptamer immobilized on the test line by "poly T"[2][3][4], the Au nanoparticles aggregates to show a colour change from colorless to red, reporting at the test line.
References
1.Jauset-Rubio M, Svobodová M, Mairal T, et al. Aptamer Lateral Flow Assays for Ultrasensitive Detection of β-Conglutin Combining Recombinase Polymerase Amplification and Tailed Primers. Anal Chem. 2016;88(21):10701-10709. doi:10.1021/acs.analchem.6b03256
2.Chao Zhu, Guilan Zhang, Yafei Huang, Shuming Yang, Shuyue Ren, Zhixian Gao, Ailiang Chen, Dual-competitive lateral flow aptasensor for detection of aflatoxin B1 in food and feedstuffs, Journal of Hazardous Materials https://doi.org/10.1016/j.jhazmat.2017.10.026
3.Wu, S., Liu, L., Duan, N. et al. A test strip for ochratoxin A based on the use of aptamer-modified fluorescence upconversion nanoparticles. Microchim Acta 185, 497 (2018). https://doi.org/10.1007/s00604-018-3022-0
4.Delley CL, Liu L, Sarhan MF, Abate AR. Combined aptamer and transcriptome sequencing of single cells. Sci Rep. 2018 Feb 13;8(1):2919. doi: 10.1038/s41598-018-21153-y. PMID: 29440771; PMCID: PMC5811598.