Team:Costa Rica/WetLab

These are the protocols that our team used in the wet lab. We present each protocol with an enumerated list of steps that we followed to accomplish each procedure. If a modification of the protocol was needed, it was specified in the notebook so everyone can follow our steps.

Escherichia coli

Lactobacillus casei

Molecular Biology

Clostridium difficile

Induction of AIP in E.coli BL21 strain

Plasmid DNA Extraction using Alkaline Lysis Method

Colony PCR

Chemically Competent E.coli DH5ɑ Cells Transformation

Growth curves of different organisms

Purification of CDL21L1-179 endolysin in E.coli BL21 strain

Quantification of bacterial fluorescence

Intracellular protein extraction from E.coli BL21 strain

Protein induction in Escherichia coli BL21 strain

Cryopreservation of different bacterial strains

1 - Tranformation of Lactobacillus casei ATCC 334TM

2 - Tranformation of Lactobacillus casei ATCC 334TM

Protein induction in Lactobacillus casei ATCC 0334

Cryopreservation of different bacterial strains

BioBricks Resuspension

Colony PCR

Dialysis and concentration of proteins

DNA Ligation (Thermo)

Lysine Susceptibility Testing on Clostridium difficile NTCC 13307

Particle Standard Curve with Microspheres

PCR

iGEM Prefix and Suffix Restriction

Polyacrylamide Gel Electrophoresis (SDS PAGE)

Fluorescence Standard Curve with Fluorescein

QS toxins induction in Clostridium difficile NTCC 13307

RNA Extraction from Clostridium difficile

"La ciencia es lo que entendemos lo suficientemente bien como para explicarle a una computadora. Arte es todo lo demás que hacemos".

— Donald Knuth.