This is a post to say that we have been implementing the cellular reagent version of the OpenVent_mCherry in Santiago (Chile) and it’s working! A big shoutout to our collaborators in Mendoza (Argentine) from Dra. Damiani’s lab (Mariangeles Avila in particular) who facilitated us the plasmid for OpenVent_mCherry.
We did have some difficulty with the induction of the expression of the polymerase though, so we wanted to share the experience. Here’s a document that systematizes what we have been doing, hopefully someone finds it useful.
Thanks to @jenny_molloy too for her guidance during her stay in Santiago a couple of months ago. We have a workshop in a couple of weeks with high school teachers in Southern Chile to produce the OpenVent locally so we are pretty excited about those results!
An update regarding the last post, just to let you know that we have been doing a workshop with high school teachers in Punta Arenas (Chile) and we produced OpenVent there. The activity was successful, we are very happy with the results because the teachers didn’t know how to use a micropipette at the beginning of the workshop and they ended up producing their own polymerase and performed successful PCR reactions. We used a lot of open-source low-cost hardware to perform the different steps and we left the teachers with a starter kit to repeat this experience in their own schools.
Then we went to Tierra del Fuego and repeated the production, we observed activity of the OpenVent that was produced in the plates we use to inoculate autoinduction media. But when trying to replate the stock in a one-week-old autoinduction media we observed very low expression. This backs up a conversation we had with @jenny_molloy regarding the relevance of using fresh autoinduction media.
Here is a document that describes the whole process, in case anyone is interested.