Open AccessSupplements
Author(s) -
R E Leach
Publication year - 1964
Publication title -
acta obstetricia et gynecologica scandinavica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.401
H-Index - 102
eISSN - 1600-0412
pISSN - 0001-6349
DOI - 10.3109/00016346409160074
Subject(s) - medicine , citation , information retrieval , library science , computer science
Supplements Materials and Methods Manipulation of X. laevis embryos Wild type X. laevis eggs were obtained by injecting 1000 IU of human chorionic gonadotrophin into the dorsal lymph sacs of adult females 6–8 hours before egg collection. Eggs were fertilized in vitro with minced testes, dejellied with 2% cysteine hydrochloride (pH 7.8–8.0) 30 min after fertilization, and cultured in 0.1X MBS (1.76 mM NaCl, 48 mM NaHCO3, 20 mM KCl, 200 mM Hepes, 16 mM Mg2SO4, 8 mM CaCl2, 6 mM Ca(NO3)2, pH 7.4) buffer. Staging of X. laevis embryos was according to Nieuwkoop and Faber [1]. Cas9 mRNA (300 pg/embryo) and gRNA(s) (each 300 pg/embryo) in 5 nl volume was injected into one-cell stage embryos from the animal pole.