Apr 12, 2022

Public workspaceMaternal Lines Protocol V.2

This document is a draft, published without a DOI.
  • 1University of Michigan - Ann Arbor
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Document Citation: Meghan Duffy, Rebecca Bilich 2022. Maternal Lines Protocol. protocols.io https://protocols.io/view/maternal-lines-protocol-b7hbrj2nVersion created by Kira Monell
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: April 12, 2022
Last Modified: April 12, 2022
Document Integer ID: 60675
Abstract
This a protocol to set up and maintain Daphnia maternal lines for clones that will be used in experiments.
Maternal Lines Protocol
By Rebecca Bilich 3/2/21
Set up adult and/or visibly pregnant Daphnia of the desired clone individually in 50 mL beakers in 30-40 mL of filtered lake water. Tap with cetyl alcohol and feed 1 mL Ankistrodesmus falcatus at 1,000,000 c/mL, 3-4 times a week.
Every other day, perform a water change:
Dump the contents of the beaker into a watch glass, rinse the beaker with filtered lake water, fill beaker with fresh filtered lake water and pipet the desired Daphnia back into the beaker.
Keep track of the number of clutches the adult female has. Remove the first two clutches of babies, keep the mother, and record the clutch on the outside of the beaker (for instance, by adding a yellow sticker after the mom’s first clutch and a red sticker after the second clutch.)
Use the babies in the third clutch as the mothers of the next maternal generation. Set them up individually and repeat the process.
If a lot of animals are dying, it’s a good idea to keep the mothers of the previous generation around for a few weeks in case replacement babies are needed.
The third or later clutches born to the third generation of moms should be suitable for use in experiments, or the third clutch born to a later generation if desired.