Nematode Synchronization: A Method to Obtain Populations of Worms in Identical Stages of Development

Published: April 30, 2023

Abstract

Source: Sarasija, S. and Norman, K. R. Measurement of Oxygen Consumption Rates in Intact Caenorhabditis elegansJ. Vis. Exp. (2019).

In this video, we show a method of producing a developmentally synchronized population of sterile adult C. elegans. The example protocol synchronizes worms to study oxygen consumption rates in live animals.

Protocol

This protocol is an excerpt from Sarasija and Norman, Measurement of Oxygen Consumption Rates in Intact Caenorhabditis elegans, J. Vis. Exp. (2019). 1. Growth and synchronization of nematode population These animals will reach the L4 larval stage after ~42 h. At this time, move the L4 larvae using a platinum pick to OP50 seeded NGM plates containing 0.5 mg/mL 5-fluoro-2’-deoxyuridine (FUdR) to prevent t…

Representative Results

Table 1: Recipes for NGM plates, M9 buffer, and bleach solution. M9 buffer (1 L) dH2O 1,000 mL NaCl 5 g KH2PO4 3 g <t…

Disclosures

The authors have nothing to disclose.

Materials

100 mm; 60 mm Petri dishes Kord-Valmark Labware Products 2900; 2901
15 mL conical tubes Corning 430791
50 mL conical tubes Corning 430829
Agar Fisher Scientific BP1423-2
Bacto peptone BD; Bacto 211677
Bacto tryptone BD; Bacto 211705
Bacto yeast extract BD; Bacto 212705
Bleach Generic
Calcium chloride dihydrate (CaCl2·2H2O) Fisher Scientific C79-500
Cholesterol Fisher Scientific C314-500
Deionized water (dH2O)
Glass Pasteur pipettes Krackeler Scientific 6-72050-900
Magnesium sulfate heptahydrate (MgSO4·7H2O) Fisher Scientific BP213-1
Potassium phosphate dibasic (K2HPO4) Fisher Scientific BP363-1
Potassium phosphate monobasic (KH2PO4) Fisher Scientific P285-500
Sodium chloride Fisher Scientific BP358-10
Sodium hydroxide (NaOH) Fisher Scientific BP359-500
Sodium phosphate dibasic anhydrous (Na2HPO4) Fisher Scientific BP332-1

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Cite This Article
Nematode Synchronization: A Method to Obtain Populations of Worms in Identical Stages of Development. J. Vis. Exp. (Pending Publication), e20045, doi: (2023).

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