BRICOBIO PLASMID TRANSFORMATION WORKSHOP

BricoBio is a DIY Bio group in Montreal, with a biosafety level 1 lab located in Helios Makerspace. WhiteFeather is an incorporating member of BricoBio, and recently participated in a plasmid transformation (with E. coli) workshop led by Kevin Chen.

IMG_0401
Cultivated E. coli colonies. @BricoBio

The two-part workshop included mixing LB with agar and an antibiotic, as well as cultivating E. coli on agar. Six of the cultivated E. coli colonies were mixed with transformation buffer solution and plasmids for antibiotic resistance and red fluorescence. A Synbiota kit was used. Our protocol was only slightly different that the Synbiota protocol. The new, antibiotic resistant (and fluorescent) bacteria solution was then applied to the agar plate with antibiotic.

13266107_10153505276030009_4368938272257619155_n
Streaking our genetically modified E. coli onto agar plates. Photo credit: Peter Chen.
IMG_0410
Adding E. coli potentially transformed with plasmids for antibiotic resistance and red fluorescence to a plate of agar with antibiotics. Successful transformation would mean E. coli growth on the antibiotics (and glowing red under UV light).

Results of the experiment: Ultimately, no bacteria grew on the plates, meaning the plasmid transformation process was unsuccessful. A variety of factors were examined. Next experiments to be done will use the CRISPR Cas-9 system.

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s