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
