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For bisulfite conversion, 200 pg of control in vitro transcribed Renilla luciferase (R-Luc) RNA was added to either 2 μg of total RNA or 200 ng of purified tRNAs and converted with sodium metabisulfite (SIGMA-ALDRICH) as previously described [ 29, 51].
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For Luciferase assays, cells were transfected either 24 h after seeding in 24-well plates or 24 h after transfecting siRNAs with 475 ng of constructs cloned in pGL3-promoter vector and 95 pg of pRL control vector using Lipofectamine2000 (Invitrogen).
Median serum concentrations of HAI-1 were 2580 and 2126 pg ml−1 for this subset of control and cancer patients compared with median urine concentrations of 666 and 2759 pg ml−1.
Briefly, 50 pg pTK-Renilla and 100 pg M50 TOPFlash (Addgene, 12456) or M51 FOPFlash (Addgene, 12457) (Veeman et al., 2003) were co-injected with 200 pg of either fezf2 or control lacZ mRNA.
Dry weight of radish was significantly improved by 100%% PG, 33 % PG, PNM and OLE treatments than that of control plants.
Gelsolin expressions in PG clones were higher than those of control cells, PN and PC10 (data not shown).
Quality assessment by Agilent Bioanalyzer shows that the sscDNA fragments generated from the 50 ng and 300 pg of HUVEC RNA positive controls were of broad distribution in length, averaging approximately 500 1000 nucleotides, suggesting efficient sscDNA synthesis (Figure 7 A, B).
As a control, 100 pg of human WT LRRK2 recombinant protein (amino acids 970 2527, Invitrogen) was loaded.
In the positive control experiment, 10 pg of rye genomic DNA were used as the template; whereas in the negative control no DNA template was used.
Chondrocyte monolayer controls released 1,100 pg of VEGF per 1 ml of medium and cartilage explant controls released 540 pg of VEGF per 1 ml of medium.
Typical sensitivity limits (mean plus 3 standard deviations of six replicate 0 pg ml−1 control wells) were 0.5 pg ml−1 hβD1, 4.5 pg ml−1 hβD2 and 5.1 pg ml−1 IL-8.
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