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Normal myeloblasts, promyelocytes and neutrophils were isolated from normal bone marrow samples using fluorescence activated cell sorting (FACS).
At the time of the diagnosis, chromosome analyses were performed on bone marrow samples using a RHG-banding technique.
RNA was extracted from peripheral blood or bone marrow samples using a TRIzol®, (Thermoscientific, MA, USA) standard protocol and quantified by the nanodrop 2000 instrument (Thermoscientific, MA, USA).
Disseminated tumour cells were found in 152 of 252 (59%) bone marrow samples using IMS and in 25 of 234 (11%) samples using ICC.
Mononuclear cells (MNCs) isolated from bone marrow samples using density gradient centrifugation were treated with IFN-γ and BCGV for 2 days.
DNA was extracted from peripheral blood or bone marrow samples using either QIAamp® DNA Blood Mini Kit (QIAGEN) or PureLink™ Genomic DNA (Invitrogen).
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The 14 bone marrow samples used in this study were obtained from healthy donors operated at the Dept of Orthopedics at "Centre Hospitalier Universitaire de Nantes".
Dual-colour Fluorescence In Situ Hybridisation (FISH) was performed on the diagnostic bone marrow sample using a t(9 22) specific BCR/ABL1 dual color, dual fusion probe (Abbott Vysis, Germany).
Total RNA was extracted from the diagnostic bone marrow sample using the Trizol LS reagent (Invitrogen, Belgium) according to the manufacturer's recommendations.
High molecular weight DNA and RNA were extracted from the bone marrow sample using 1 ml of Tripure isolation reagent (Roche Diagnostics, Indianapolis, USA), according to the manufacturer's instructions.
Since the bone marrow sample used to generate the LCDD-02 sequence was taken post-stem cell transplant, we think it was presumably polyclonal.
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