P1 Lab Activities
The P1 lab is official at Hackuarium! (as of 1nov17)
We can official select for bacterial clones carrying plasmids we want 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! ;)
(If someone would like to help for translations to French or other languages, please do feel free!)
Inspiration
Many Hackuarium members have been working towards this goal since the foundation of this association three years ago!
In particular Yann P, Sam S and others did great leg-work, and even tried a 'green house' enclosure in the old space...
(Prior to renovation.)
All the work leading up to us finally getting the lab up and running is documented (sorry some bits are still redundant and in need of edits) in this older page.
The How To Grow Almost Anything course, which Dan Hernandez and Thomas Jordon really wanted to do, was further inspiration for finally getting the lab running.
Now (nov 2017 - present), Rachel Aronoff is acting as the biosafety officer for the space, and Olivier Emery has agreed to take on the role of deputy biosafety officer.
Six people have already taken the 'intro to biosafety' training, and the next training session is planned for 10nov17.
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!
DH5alpha test begun 16oct, first test streak on 2nov...
Bioluminescent strain improvement
Mixed biosensor strain (arsenic inducible GFP and constitutive mCherry) separation
List of Potential P1 Experiments
HTGAA list
Open Insulin?
others??
Key Aims
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!
Gallery
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 biolum 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.)
Further information
- 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!!