RCA Protocol: Fresh Frozen Tissue¶
This page is retired — it has been split in two
Up to v5.4 this page carried the whole fresh-frozen workflow. Its contents were verbatim duplicates of two pages that are now the source of truth:
- Sections 1--4 (sectioning, fixation, permeabilization, dehydration) → Sample Pre-treatment: Fresh Frozen
- Sections 5--9 (blocking, hybridization, ligation, RCA, post-fix) → RCA Protocol (the unified protocol, shared by all sample types)
Nothing was lost — the only fresh-frozen-specific detail, the lower ~5% glycerol in the RCA mix, is now a note in RCA Protocol §3.
Fresh frozen workflow: Pre-treatment (FF) → RCA Protocol → signal readout.
Why it was retired¶
Keeping a combined page meant every shared change — a wash duration, a probe
concentration, the RCA primer — had to be made twice, in this page and in the unified
protocol, and kept in sync by hand. It drifted: by the time it was retired this page and
rca.md disagreed on the RCA table (one summed to 100 µl, the other to 97.5 µl), and this
page had fallen out of the site navigation while still being linked from elsewhere.
The pre-treatment / library-construction split is the structure the rest of the site already uses — FFPE, cDNA and iLock all follow it.
Changelog¶
- retired -- 2026-07-30 -- Content split to pretreatment-ff.md and rca.md; FF-specific glycerol note folded into the latter. Page kept as a pointer so existing links and bookmarks do not 404.
- v5.4 -- 2026-07-29 -- Final combined release (see git history for the full text).
- v5.3 -- 2026-01-29 -- Prior fresh-frozen release.