Skills
Restriction Cloning
Restriction maps, virtual digests (fragment sizes for linear or circular DNA) and picking enzymes for restriction cloning - single cutters in a vector/MCS that don't cut the insert, directional enzyme pairs, and Type IIS (BsaI, BsmBI, BbsI, SapI) sites for Golden Gate domestication. Use with plasmid/insert sequences, FASTA or GenBank files, or questions like "what will my diagnostic digest look like?"
Qpcr Analysis
Analyze RT-qPCR Ct data for relative expression - 2^-ddCt or efficiency-corrected (Pfaffl/qbase) fold change with one or more reference genes, technical-replicate checks, Undetermined wells, group summaries, and primer efficiency from a standard curve. Use when someone shares Ct values, a qPCR export or a standard-curve dilution series.
Primer Qc
Check PCR/qPCR primers and oligos - melting temperature (SantaLucia nearest-neighbor, Primer3-style salt correction), GC%, 3' GC clamp, hairpins, self- and cross-dimers, Tm mismatch and a suggested annealing temperature. Use when someone pastes primer sequences, asks for an oligo's Tm, or asks whether a primer pair will work.