I’ve open-sourced 14 years of lab protocols and accumulated knowledge at finkelsteinlab.org/wiki.
Where to start
If you are coming from outside the lab, these are the pages most likely to be useful:
- Protein purification. Some useful starting points: CRISPR nucleases (Cas9, Cas12a, Cas13d, Cascade, Cas3), DNA repair and replication proteins (human and yeast RPA, Exo1, Dna2, BLM, MRN, Rad51, the T4 replisome), human and yeast histones, and the workhorse proteases (TEV, SUMO, sortase).
- Single-molecule imaging. Building DNA curtains and ssDNA curtains, flowcells and their troubleshooting, and particle tracking.
- Microfabrication. UV and e-beam lithography for curtain tracks, slide fabrication and dicing.
- Biochemical assays. EMSAs, cleavage reactions, ATPase and filter-binding assays, plus NucleaSeq and CHAMP.
- Protein labeling. Sortase reactions, biotinylation checks, epitope tags and quantum-dot conjugation.
- DNA, RNA and cloning and fission yeast. The everyday molecular biology, including crRNA transcription and S. pombe genetics.
- Recipes and buffers. Media, antibiotics, gel buffers, loading dyes, imaging buffers.
- Computing and code. Fitting data to a model, an introduction to diffusion, ImageJ macros and an ND2 file reader.
- Careers and writing. Writing advice, funding, and Heilmeier’s catechism for anyone pitching a project. Future writing advice will be on my blog.
The home page links every section, and the search box at the top covers the whole site.
Pull requests welcome
If you find a mistake or want to help us improve this resource, open a pull request or an issue on finkelsteinlab/wiki (every page has an Edit page link). And if a protocol from one of our papers is not on the wiki yet, ask, and we’ll try to add it in due time.