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This project, funded by ANID-CONCYTEC CovBio, aims to develop low cost and open diagnostics for SARS-CoV-2 based on two main isothermal reactions: RT-LAMP and RPA. This repository will collect data and release up-to-date progress. #### **Preliminary protocols for enzyme purification for RT, RPA and LAMP** * **BstLF**: [Expression and purification protocol][1], [genbank seq][2] and [discussion thread at ReClone][3] * **MMLV RT**: [Expression and purification protocol][4], [genbank seq][5] y [discussion thread at ReClone][6] * **HIV RT**: https://dx.doi.org/10.17504/protocols.io.e6nvwje79lmk/v1 * **Bst-LF DNA pol**: http://dx.doi.org/10.17504/protocols.io.bksrkwd6 * **MMLV**: http://dx.doi.org/10.17504/protocols.io.bsernbd6 * **Taq Pol**: http://dx.doi.org/10.17504/protocols.io.bya3psgn * **Pfu-Sso7d**: http://dx.doi.org/10.17504/protocols.io.bzusp6we * **Bsu**: dx.doi.org/10.17504/protocols.io.x54v9ypkpg3e/v1 * **uvsX**: dx.doi.org/10.17504/protocols.io.5qpvobemdl4o/v1 * **uvsY**: dx.doi.org/10.17504/protocols.io.36wgq7xwkvk5/v1 * **gp32**: dx.doi.org/10.17504/protocols.io.kxygx957wg8j/v1 * **LbCas12a**: https://www.protocols.io/private/0766389C528AAD67EC807C8A06D8516B #### **Preliminary results** * **RT:LAMP**: [Buffers and reactions were prepared according to these open protocols][7] and [Code for reaction optimization][8]. * [Manuscript][9] and [data repo link][10] * [Open notes about RPA progress][13] Meeting notes can be found [here][11] All the DNAs are accessible under [openMTA][12] [1]: https://dx.doi.org/10.17504/protocols.io.bksrkwd6 [2]: https://benchling.com/s/seq-MuNUtRESOLQyYjeRVm8Z [3]: https://forum.reclone.org/t/bst-large-fragment/91/10 [4]: https://www.protocols.io/view/recombinant-expression-and-purification-of-codon-o-bhcvj2w6 [5]: https://benchling.com/s/seq-LZOeOlZ90svPCI5vrlQF [6]: https://forum.reclone.org/t/mmlv-reverse-transcriptase/88 [7]: https://www.protocols.io/edit/low-cost-lamp-and-rt-lamp-bsejnbcn [8]: https://github.com/LabTecLibres/RT-LAMP [9]: https://abrf.memberclicks.net/assets/JBT/September_2021_Early_Access/Homebrew%20reagents%20for%20low-cost%20RT-LAMP.pdf [10]: https://zenodo.org/record/4540817#.YXbgGNlKhTZ [11]: https://docs.google.com/document/d/1TTkAaMQV8mwUSLunZ28Ag2lu80zlN-QHa7FgEX0LMdk/edit?usp=sharing [12]: https://biobricks.org/openmta/ [13]:https://docs.google.com/document/d/1U0hOLi_fPEkSS2fFpEe7t3jbY_xZjOj0/edit?usp=sharing&ouid=108512776541224231501&rtpof=true&sd=true
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