Tag:
Genetic Engineering
Biologics (antibody+ADC)
McClean: Working with the LoxP/Cre System in S. cerevisiae-PDF
Overview
The loxP/Cre recombinase system is an extremely powerful tool for genetic manipulation, allowing for conditional and site-specific integrations, excisions, and inversions, either in vivo...
gene therapy & gene editing
Site-directed mutagenesis/Single site-PDF
Materials
Template plasmid purified from a dam+ E. coli strain
Mutatagenesis primers
PfuTurbo DNA polymerase (nonstrand-displacing) and associated reaction buffer
dNTPs
PCR Thermocycler
Dpn...
assay development
Electroporation-PDF
This protocol is for transforming plasmid DNA into Escherichia coli cells.
Materials
Electrocompetent cells
Plasmid DNA (from a ligation reaction)
Ice
Ice bucket
For the following,...
assay development
Lactobacillus chromosomal integration-PDF
Overview
This procedure is used to integrate a desired DNA cassette into the chromosome of Lactobacillus plantarum at specific locations. This protocol can also be...
Biologics (antibody+ADC)
High Efficiency Transformation-PDF
Protocol
Day 0
Make sure you have the necessary solutions (instructions for how to make them can be found here):
Single-stranded carrier DNA
PEG 3350...
assay development
McClean: Designing “Yeast Toolkit” compatible primers-PDF
Overview
This protocol is written to prepare the user to design PCR primers such that the end product will have BsaI and BsmBI cut sites,...