Experiments
Experiment design¶
- Workflow for performing experiments
- Labeling conventions for experiments and samples
- Conventions for storing plasmids, strains, and cell lines
- Tips for minimizing errors in molecular biology experiments
Buffers¶
- Lithium acetate borate fuffer for agarose gel electrophoresis
- RNA extraction buffer
- SDS PAGE buffer (Tris-Glycine)
- TBE buffer for RNA gel electrophoresis
- Transfer buffers for Western blots
- TBS(T) buffer
Biochemistry¶
RNA¶
Protein¶
- Extracting proteins from TRIzol lysate flowthrough
- Standard Western blot protocol (to be added soon)
- Validated antibodies for Western blots
Links¶
- Accessible introduction to Protein Chromatography using AKTA
- Size Exclusion Chromatography – Principles & Methods
Cloning¶
- Basic Phusion PCR protocol
- Choosing restriction enzymes to verify cloning
- Cas9-mediated Genome Editing in S. cerevisiae
- Designing primers for Gibson cloning
- How to mutate using BsmB1-mediated Golden Gate assembly
- How to design on- and off-target sgRNAs for CRISPR-mediated genome editing
- Large scale yeast chemical transformation
- Preparing Gibson isothermal assembly mix
- Preparing chemically competent E. coli
- Preparing chemically competent E. coli with one-step TSS
Links¶
- NEB Golden Gate Assembly Tool This is useful for assembling repeat regions with linkers such as in 24xMS2 or 24xHA. How to: https://www.neb.com/golden-gate/using-the-neb-golden-gate-assembly-tool. Tool link