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Oocytes and sperm were obtained by dissection of gonoducts, dechorionation was performed with 1.0% sodium thioglycolate (T0632, Sigma) and 0.05% actinase E (D4527, Sigma) (Mita-Miyazawa et al., 1985) and cross fertilization was performed in glass tubes.
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Then artificial fertilization was performed in the same manner as in trial 1.
Oocyte fertilization was performed by ICSI.
Fertilization was performed under previously described standardized conditions [ 16].
Artificial fertilization was performed following [ 45] with some modifications.
Fertilization was performed by transferring conidia from the mat a strain (8782) to the Petri dish containing maternal tissue of the mat A strain (8882), and vice versa, to produce two reciprocal crosses.
A cross-fertilization experiment was performed at Lautiosaari State Fish Hatchery, Keminmaa, Finland in 1993 94.
Three independent biological replicates of the in vivo cross-fertilization assays were performed with highly consistent results, each using a different mouse and mosquito batch.
Fertilization of oocytes was performed using classical in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI).
Fertilization by Hoagland solution was performed once a week.
The biopsy procedure was performed three days after fertilization.
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