Week 2 of Pipetting adventures

Shout out to the TAs for their time and patience.

Spectinomycin is an antibiotic that inhibits bacterial protein synthesis by binding to a ribosomal subunit. Bacteria have resistance to spectinomycin by enzymes that inactivate the antibiotic by covalently attaching adenosine monophosphate molecule to the amino acid side chain.

Step I : Restriction Digest to generate a ladder

using PstI and EcoRV to cut plasmids pPSU1 and pPSU2

First, I used benchling to find cut sites for each restriction enzyme on pSU1 and pSU2 seperately.

pSU1 and EcoRV-HF cut sites on Benchling

pSU1 and PstI-HF cut sites on Benchling

pSU2 and EcoRV-HF cut sites on Benchling

pSU2 and PstI-HF cut sites on Benchling

Gel Electrophoresis in the lab after restriction enzyme reaction

To find the EcoRV-HF/PstI-HF restriction enzyme products of plasmids pSU1 and pSU2, we had to mini prep the plasmids from the provided pSU1 and pSU2 cells.

Step 1a - Rapid and small-scale isolation of plasmid DNA from bacteria using Qiagen Miniprep Kit

Protocol