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Sf21 cells (Invitrogen) and Sf21-gfp reporter cell line were cultured in TNM-FH Insect Medium (BD BaculoGold) supplemented with Grace's medium including trace metals, lactalbumin hydrolysate, yeastolate, and 10% heat inactivated fetal bovine serum along with 300 μg/ml Zeocin (100 mg/mL) (Invitrogen).
Schneider for insect medium was purchased from Sigma-Aldrich (St . Louis MO, USA).
Wing discs were dissected from 3rd instar larvae in Shields and Sang M3 Insect Medium (Sigma) without serum.
Drosophila cultured cells were maintained at 24°C in Schneider's insect medium (Gibco) supplemented with 10% fetal bovine serum (JHR) and antibiotics (Invitrogen).
Drosophila larvae were secured to a glass dish in 2 ml of Schneider's Insect Medium (Sigma) using a blunt magnetic placed pin distal to the CNS.
Promastigote forms were maintained by weekly transfers in 25-cm2 culture flasks with Schneider's insect medium, pH 7.0, supplemented with 10% FBS at 26°C.
The growth medium consisted of equal volumes of Mitsuhashi-Maramorosch (MM) and Schneider's Insect medium (Sigma) supplemented with 20% heat-inactivated fetal bovine serum (Sigma).
Briefly, late 3rd instar larvae at wandering stage were rinsed in ice-cold Schneider's insect medium (Sigma) supplemented with complete Mini Protease Inhibitor Cocktail (Sigma).
Dissected tissue was rinsed and cut into segments in sterile Grace's Insect Medium (Gibco BRL, # 11590-056) and transferred into 24-well culture plates.
Salivary glands and imaginal discs from wild type and Top2f third instar larvae were dissected and placed into serum free HyClone CCM-3 Insect medium.
Drosophila S2 cells were cultured at 24°C in Schneider's insect medium (Sigma) supplemented with 10% heat-inactivated fetal bovine serum.
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