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Embryo Extract:

1.    Chill 200, 3 d embryos after removing from chorions.
2.    Rinse in 0.5% chilled bleach for 2 minutes and then in zero calcium Ringer for 2 minutes.
3.    Transfer to a Dounce homogenizer with a minimum of liquid and homogenize well.
4.    Resuspend in 1 ml L-15 supplemented with 0.3 mg/ml glutamine, 50 U/ml penicillin, 0.05 mg/ml streptomycin and 0.8 mM CaCl2.
5.    Store at -20°C.

Growth medium:

L-15 (Sigma)

0.3 mg/ml glutamine

50 U/ml penicillin

0.05 mg/ml streptomycin

0.8 mM CaCl2

10% embryo extract

3% fetal calf serum.

Filter through a syringe filter.

1 Comment

  1. Jul 13, 2012

    Anonymous

    Hi, Just a precision I need for the growth medium : is it L-15 with or without …

    Hi,

    Just a precision I need for the growth medium : is it L-15 with or without glutamine from Sigma ?  Indeed, the labelling is similar in their catalog

    Thanks for answering,