Scale alkaline hypochlorite reagents for any number of gravid plates and predict L1 → young adult stage pickups at 15, 20, or 25 °C.
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Load example celegans synchronization calculator data to see the full workflow
Each plate uses 4 mL water + 1.5 mL bleach + 0.5 mL 5N NaOH (6 mL total).
Standard rearing — Q10≈2 baseline.
If provided, the calculator returns ETAs for each stage transition.
| Reagent | Volume |
|---|---|
| Distilled water | 22 mL |
| Sodium hypochlorite (5–6% bleach) | 8.3 mL |
| 5 N NaOH | 2.8 mL |
| Stage | Hours after start | ETA |
|---|---|---|
| L1 (hatched in M9) | 14 h | — |
| L2 | 26 h | — |
| L3 | 34 h | — |
| L4 | 42 h | — |
| Young adult (gravid) | 54 h | — |
When to use
Do not use for
Chlorine activity in alkaline hypochlorite drops within minutes. Mix water + bleach + NaOH right before you add it to the worm pellet, and never store leftover solution.
Bleach for too short and you get adult debris in your egg prep. Bleach for too long and you damage the eggshells. The right endpoint is "no intact adults visible" — usually 5–8 min.
Three M9 washes is the minimum. Residual bleach kills L1s during the overnight hatch. If your hatching efficiency is low, add a fourth wash.
Temperature predictions are guides, not guarantees. Score 5–10 worms under the dissecting scope before committing to a large pick — vulval crescent for L4, embryos at spermatheca for gravid YA.
Reagents: per-plate volumes (4 mL water + 1.5 mL bleach + 0.5 mL 5N NaOH) plate count 1.1 overage. Stage timeline: cumulative 20 °C intervals (L1@14 h, L2@+12 h, L3@+8 h, L4@+8 h, YA@+12 h) scaled by Q10≈2 temperature factors (15 °C ×1.6, 20 °C ×1.0, 25 °C ×0.65). ETAs computed by adding scaled hours to the user-provided bleach start datetime.
Last validated 2026-04-06. Calculations are designed for planning and documentation support; verify procurement decisions against manufacturer specifications or institutional SOPs.
ConductScience C. elegans Bleach Synchronization Timing Calculator (v0.97.0). ConductScience, Inc. 2026. Available at: https://conductscience.com/tools/celegans-synchronization-calculator
Stiernagle T. Maintenance of C. elegans. WormBook. 2006. doi/10.1895/wormbook.1.101.1
Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974;77(1):71-94.
Adult C. elegans cuticles dissolve in alkaline hypochlorite within 5–8 minutes, but eggshells (chitin) survive intact. This gives you a clean cohort of eggs that hatch synchronously as L1s — the foundation of any age-matched experiment.
Always make this fresh — the chlorine activity drops fast once NaOH is added.
1. Wash gravid plates into a 15 mL conical with M9 buffer 2. Spin at 1,500 g for 1 min, aspirate supernatant 3. Add 6 mL fresh bleach solution, vortex briefly 4. Watch under a scope — adults dissolve in 5–8 min, leaving eggs intact 5. Spin, aspirate, wash 3× with sterile M9 6. Resuspend in 5 mL M9 in a fresh conical and rotate overnight at 20 °C 7. Eggs hatch as starvation-arrested L1s in 12–16 hours
After hatching as L1 in M9, the cohort develops on OP50-seeded NGM plates with predictable timing. Use the calculator to schedule pickups.
Keep eggs in M9 with no food for 12–16 h at 20 °C. Worms hatch and arrest at L1 — the most synchronized timepoint you can get. Plate to OP50 to release the arrest and start development.
At 20 °C, L4s appear ~42 h after bleach start. The vulval crescent is unmistakable under a dissecting scope. Pick L4s for: lifespan setup, RNAi feeding plates, synchronized aging cohorts, mating crosses.
At 20 °C, gravid young adults appear ~54 h after bleach start. Pick YAs before egg laying begins (no embryos in the gonad) for: brood size assays, fecundity scoring, single-worm sorting.
Schedules bleach synchronizations across your C. elegans colony, sends stage pickup alerts, and tracks every cohort from egg to assay endpoint.
Open app EquipmentM9 buffer, sodium hypochlorite, NaOH, and culture supplies for bleach synchronization
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