P1 Lab Activities
A P1 laboratory was officially functional at Hackuarium between 1 November and 1 September 2018. The Hackuarium community laboratory had to move to another location at this point.
This allowed members to select for bacterial clones carrying plasmids they wanted to use or make!
'Construct' is the proper noun for a plasmid, and 'to construct' is the proper verb for this process, btw. Even if many people like to talk about cloning.
This page is still lacking a French translation. If you are interested in helping, please reach out!
Several Hackuarium members have been working towards the goal of having a certified P1 laboratory since the foundation of the association in the spring of 2014.
In particular Yann P, Sam S and others did great leg-work, and even installed a 'green house' enclosure in the former open space (prior to renovation of UniverCité's second floor space).
All the work leading up to us finally getting the lab up and running is documented in this page (work in progress).
One catalyst to get the laboratory equipped was the participation of Dan Hernandez and Thomas Jordon to the "| How To Grow Almost Anything" (HTGAA) course organised by MIT professor George Church.
One aspect that was particularly important to Hackuarium was to have professionally trained biosafety officers to oversee the work done in the P1 laboratory.
Rachel Aronoff is acting as the biosafety officer for the space, while Olivier Emery has agreed to take on the role of deputy biosafety officer.
If you need to contact them in particular about any proposed or in progress P1 activities, your can contact them via this new email address: firstname.lastname@example.org
Eight people have already taken the "Introduction to biosafety" training. More training sessions can be organised upon request.
P1 experiments at Hackuarium
Initially, these will all be limited to bacterial growth of plasmid constructs.
Both classical and 'synthetic biology' strategies are possible!
List of P1 Experiments
Make sure to link your project pages here!
(more to come soon! )
DH5alpha test begun 16oct, first test streak on 2nov...
Bioluminescent strain growth
Mixed biosensor strain (containing 2 plasmids, an arsenic inducible GFP and constitutive mCherry)
List of Potential P1 Experiments
HTGAA set coming soon!
To learn, have fun, and avoid problems!
Here is the link to all the P1 safety info, list of users and list of strains for future reference.
Watch this space for more soon!
The first images are of plates carrying our first plasmid strains, streaked at UNIL and grown in our own incubator, and of Vlad streaking the bioluminescent strain...
The big -80 freezer behind him can hold our stock, at DMF UNIL, until we know if the -20 method will work, or are more fully equipped!
To note: The bioluminescent strain is supposed to be moderately expressed, but on the plate after overnight incubation at 37oC, almost no light is visible by eye...
A liquid culture will be attempted next...
Vlad helpfully found it in the DMF collection, but more work is planned to make it bright!
The second strain actually carries two plasmids, a constitutive mCherry one, and an arsenic inducible GFP one... The experiment being tested is whether, by plating this strain on the two different antibiotics, new sub-strains can be selected carrying each plasmid alone... (we already have stocks of the parent strain in the UNIL freezer! It was actually made by Rachel for her 'nematode assisted biosensor analyses' from a few years back!)
There are a couple more images for the test of bacterial strain stability in the -20 freezer, with the DH5alpha strain frozen in either glycerol vs milk. This isn't strictly a P1 experiment, because these bacteria were not selected and carry no plasmids... If they are stable, however, in other words, if they can be repeatedly propagated from this stock, this will mean our P1 lab may not need (even though it would be good if we were able to get one!) a -80 freezer, as is the usual standard for strain stocks...
Come join us!
Timeline for Project Initiation
1) Talk with everyone about your ideas and what you would like to do.
2) Take the safety course and do risk assessment on your idea.
3) Start documenting your idea in the wiki. You can use this template developed by YannP as a guide.
4) Fill in the form, also developed by YannP!
5) Get going! (ask for a minigrant, if necessary for obtaining reagents or other materials.)
- Here is the P1 standard operating procedure, which is still a work in progress!
See something that needs mentioning? You can help contribute!
- Do you have questions? Maybe you can't reach Rachel or Olivier, or want to find out more before even trying to ask them?? There is a service in the DIYBio community for this Ask a Biosafety Professional! Use it!
Remember, being clueless is no excuse!!
- The sink has now (as of 15nov17) been hooked up, but ventilation of the fume hood is (so far) only promised...