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Before hybridisation, samples were cut to 5 μm sections, deparaffinised and pretreated with commercial pretreatment kit Vysis (Abbott Molecular, Des Plaines, IL, USA).
Air-dried tissue sections were treated with a Paraffin Pretreatment Kit (Vysis Inc).
Tissue sections were treated using Paraffin Pretreatment kit II (Vysis, Downer's Grove, IL, USA) according to the manufacturer's instructions.
Paraffin-embedded LCLC sections (4 µm) were baked, deparaffined by washing in xylene, and dehydrated in 100% ethanol, and then incubated in pretreatment liquid (paraffin pretreatment kit) for 20 to 25 min at 80°C.
EGFR gene status evaluation was performed on 3 μm thick tissue sections that were treated using Paraffin Pretreatment kit II (Vysis, Downer's Grove, IL, USA) according to the manufacturer's instructions.
Slides were then treated with 2 N HCl for 20 min, Vysis Pretreatment Kit I (Abbott Molecular), washed with a 2 × SSC buffer, and incubated in pretreatment buffer at 80 °C for 30 min.
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Sections were pretreated using the Paraffin Pretreatment Reagent Kit II (Vysis): the slides were placed in the pretreatment solution at 80°C for 50 70 min and in pepsin solution 0.05% at 37°C for 20 25 min.
Fluorescence in situ hybridisation on paraffin-embedded breast tumour samples was performed using a modified Paraffin Pretreatment Reagent kit protocol (Vysis) as described previously (Rauta et al, 2006).
For proteolytic slide pretreatment, a commercial kit was used (paraffin pretreatment reagent kit; Abbott, Wiesbaden, Germany).
Sections were deparaffinized in xylene, hydrated through a graded ethanol series, and then incubated with the accessory kit pretreatment solution at 95 100°C for 10 minutes.
A commercial kit (paraffin pretreatment reagent kit 1, Abbott Laboratories, IL, USA) was used for proteolytic slide pretreatment.
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