Experiments

Mad science with the Bean Team

Constructing, Introducing, Screening, Replicating, and Isolating the Newly Constructed Plasmid

Newly constructed plasmids are integrated into E. coli as a means to screen correctly assembled colonies, replicate it into quantifiable amounts, and then isolate it back out to be used for the next assembly.


Restriction Enzyme Sequence Verification

While the antibiotic resistance and blue/white coloration suggest a successful goldengate assembly, this can be further verified through a restriction enzyme digest and gel electrophoresis.


Competent Cell Preparation

It is much cheaper to reproduce your own stocks of electrocompetent cells than to be consistently buying more. Competent cell preparation consists of multiplying a strain of cells in a nutrient rich broth, spinning them down, and rinsing them several times to remove any salts present. These new stocks of cells are then aliquoted into eppendorfs and stored at -80*C.