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In the AC protein comparison experiment, AC fluid from 14 patients was digested, analyzed by LC-MS/MS using a semi-quantitative label-free approach and the data were compared by principal component analysis (PCA) to gain knowledge of protein homogeneity of AC.
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RT-PCR products from lung cancer patients were digested with λ-Exo and analyzed using electrochemical biosensors.
In addition, tumor fluid and frozen tumor fluid from two patients were digested by trypsin and analyzed by LC-MS/MS in triplicate without any prior separation.
Briefly, adipose tissue from non-oncogenic patients were digested with collagenase P and cultured (37°C, 5% CO2) in MSC medium (DMEM plus 10% FCS, 2 mM glutamine, 50 µg/ml gentamycin) and passaged when they reached 85% confluence.
Tissue samples from central neurocytoma and oligodendroglioma patients were digested with proteinase K at 56°C overnight, and genomic DNA was isolated by phenol-chloroform extraction using a commercially available kit according to the manufacturer's procedures.
Tissues from the endometrium of each patient were digested with RNase-free DNase I (Ambion) to remove genomic DNA.
In a series of 10 experiments using samples from different patients, OA synovium was digested as specified above.
According to methods described by Doherty et al (2002), this tissue from a patient with TiVaAl prosthesis was digested in 1% trypsin-collagenase gently mixed and incubated at 37°C for 24 h.
Tissue collected from a consented patient undergoing a voluntary reduction mammoplasty was digested with 2 mg/mL of collagenase (Sigma) and 100 U/mL of hyaluronidase (Sigma) at 37°C overnight to generate organoids.
Lane 2: pXba was digested by DraIII.
Total RNA was digested with DNase I™.
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