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To confirm the expression of established markers of endochondral ossification found in our array to the literature, we first examined our microarray data sets for the expression of early stage chondrocyte markers such as Sox family members 5,6 and 9 (Sox5,6 and 9), Col2a1, growth differentiation factor 5 (Gdf5), Agc 1, Col11a1, Hapln1, Fgfr3 and Col9a2 (Figure 1B, top panel).
We selected the top 20 precursors for MS2 analysis which consisted of higher energy collision dissociation (HCD) with the following parameters: resolution 17,500, AGC 1 × 10, maximum injection time 60 ms, isolation window 2 Th, normalized collision energy 25, and centroid spectrum data type.
Orbitrap spectra (AGC 1 × 106) were collected from 400 2000 m/z at a resolution of 60 k followed by data-dependent HCD MS/MS (collision energy 32%, AGC 5 × 10) of the ten most abundant ions, excluding single charge states.
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The total noise power σ noise 2 is thus higher and includes both the contribution due to thermal noise σ thermal 2 and the AGC noise σ AGC 2 : σ noise 2 = σ thermal 2 + σ AGC 2 (14).
Both the thermal noise power σ thermal 2 and AGC noise power σ AGC 2 contribute to the total noise power σ noise 2 as indicated by the dashed curve.
The scan sequence began with an MS1 spectrum (Orbitrap analysis, resolution 60,000, 300−1500 Th, AGC target 1 × 10, maximum injection time 150 ms).
Among the di- and tri-nucleotide repeats, AC/TG (57%) and AGC (12%) were the most abundant type.
We used the relationship between belowground carbon (BGC) and AGC from the FIA data to develop a BGC spatial distribution at the same scale as AGC [5, 27].
Within accordance with NERC [22], the energy-reserve for a certain event (contingencies/outage elements interconnected) is classified by contingency reserve (AGC, R10, R30), i.e. instantaneous or ramping reserve (build-up or non-instantaneous).
The operation of the reserve system (AGC, R10, R30) is characterized by its speed of response (starting time and ramp rate), duration of response, frequency of use, use direction (up or down), and type of control.
Viral RNA was extracted using Trizol reagents (GIBCO/BRL) and reverse transcription was performed using oligonucleotide influenza universal primer Uni12: 5'-AGC AAA AGC AGG-3' [43].
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