thethoughtemporium

How I defeated my arch nemesis: BL21 E. coli (Patreon)

Published:

2021-02-23 00:54:01

Imported:

2026-06

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Bl21 ecoli are a biolgists nightmare. They are the best strain for producing proteins in ecoli, but are maddeningly difficult to get DNA in to. After a couple years of tinkering with the protocol, I've finally landed on one that works incredibly well and felt I needed to share it so others don't need to suffer through dozens of failed transformations.

Releasing this early since I sent it to the sponsor to approval. Once I get that approval it will go live, but if they have issues I'll need to reupload it with the changes. So keep that in mind if you comment. I love your comments, so maybe just save them till it goes up officially so they don't get deleted if I reupload.

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How I Defeated My Arch Nemisis: BL21 E. coli

For NordVPN’s 9th birthday, every purchase of a 2-year plan will get you 1 additional month free AND a surprise gift. Go to https://www.nordvpn.com/thoughtemporium and use code thoughtemporium! ________________________________________________________________________ BL21 E. coli are by far the most obnoxious bacteria genetic engineers usually work with for 1 simple reason. They are maddeningly difficult to modify and load with new DNA. This wouldn't be an issue if they weren't also one of the best types for producing proteins en mass, and as such have application in everything from pharmaceutical production, to producing ingredients for foods. But after a few years of tinkering with it I finally found a protocol that reliably fixes this irritating issue, and makes it easy to transform them reliably. Written protocol - https://github.com/thethoughtemporium/Protocols/blob/main/Transformations/bl21%20transformation.md Previous videos; Modifying Bacteria - https://youtu.be/F0_q-fD_lyU Modifying yeast - https://youtu.be/DHNPnO5UOYQ Making a heat block/incubator - https://youtu.be/2hf9yN-oBV4 Growing spider silk -https://youtu.be/2hf9yN-oBV4