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For the MAQC samples, 100 pg total RNA was amplified for 23 cycles and the amplification product was again analyzed on Gene ST microarrays.
We therefore proceeded with this methodology, yielding on average 8 pg total RNA per HUVEK cell, for downstream microarray probe generation workflows, to achieve global transcriptomics from input RNA in the low picogram range.
Additionally, the WG-DASL assay yielded high self-correlations (R2>0.98) with low intact RNA inputs ranging from 1 ng to 100 ng; reproducible expression profiles were also obtained with 250 pg total RNA (R2∼0.92), with ∼71% of the probes detected in 100 ng total RNA also detected at the 250 pg level.
Typical yields were 1 pg total RNA/cell.
Additional information about the PG total sample characteristics are detailed in the Appendix.
The detection limit for the pan-AAV primers PCR system was 100 pg total HA-16 DNA.
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Similarly, IL-6 protein was not detected in mucosal tissue samples from healthy controls and IL-1 β, IL-8, and TNF- α were only measured at low concentrations (median concentrations; IL-1 β 2.6 pg mg−1 total protein, IL-8 0.2 pg mg−1 total protein, TNF- α 0.1 pg mg−1 total protein).
Renin-mRNA concentration was quantitated in the kidney (1.74 +/- 0.2 pg renin/micrograms total RNA), adrenal gland (1.15 +/- 0.15 pg renin/micrograms total RNA), placenta (0.7 +/- 0.1 pg renin/micrograms total RNA), and saphenous vein (0.02 +/- 0.01 pg renin/micrograms total RNA).
The small amount of RNA within a mammalian cell (approximately 20 40 pg of total RNA [11]) presents a challenge for single-cell transcriptome analysis.
Complete SNP profiles were obtained from 10 pg of total DNA.
Some of the primers (LCVCPF1/LCVCPR1, LCVREPF2/LCVREPR2, LCrep1F/LCrep2R) could detect the virus in 2.4 0.24 pg of total DNA of infected plant.
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