Recipe

Embryo Collecting Buffer I

In advance, prepare a 7× stock solution of complete, Mini, EDTA-free Protease Inhibitor Cocktail (Sigma-Aldrich 11836170001) and a 100 mm stock solution of spermine (e.g., Sigma-Aldrich S1141-1G) in DNase-, RNase-free H2O. Store both at −20°C until use. Prepare the recipe as follows:

2× Stock Solution I 400 µL
1 m DTT (e.g., Sigma-Aldrich 646563, once opened, store at −20°C) 1.6 µL
7× cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail 114.4 µL
100 mm Spermine 0.8 µL
Protector RNase inhibitor (Sigma-Aldrich 3335399001) (40 U/µL) 4 µL
80% (w/v) glycerol (made from e.g., Sigma-Aldrich G5516-500ML) 279.2 µL

  • Prepare fresh in a 1.5-mL microcentrifuge tube (RNase/DNase-free) and store on ice. This amount of buffer is appropriate for collecting 72 embryos.

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  1. doi:10.1101/pdb.rec107965 Cold Spring Harb Protoc 2022: pdb.rec107965- © 2022 Cold Spring Harbor Laboratory Press
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