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The leaf was used to monitor expression of the three cloned genes.
These constructs were used in different vector backgrounds in which the presence or absence of GFP allowed us to monitor expression of the shRNAs (see MATERIALS AND METHODS).
The second construct is a reporter mRNA which consists of eight binding sites (19-base stem-loop) for the MS2 coat protein, a ß-galactosidase coding sequence to monitor expression of the construct and the SV40 3'UTR (Untranslated Region) to mimic the behaviour of non-targeted mRNAs.
In our study, a single viral vector was engineered containing all relevant information to induce and monitor expression of the target gene in vivo, thus reflecting the situation which might be applied in a diagnostic or therapeutic paradigm for clinical application.
A biological sample corresponding to the A. thaliana/ P. parasitica interaction was added, to monitor expression of the selected sequences during the necrotrophic stage.
To monitor expression of the hIP by indirect immunofluorescence, HEK.hIP cells were grown to 60 70% confluency on poly- l-lysine pre-treated coverslips in 6-well plates.
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Following systematic knock-down of LIN28, LIN28B and MYCN we monitored expression of the Let-7 alleles.
As another indicator of auxin redistribution in induced SS12K roots, we monitored expression of the auxin responsive reporter DR5 GFP [25] in the absence and in the presence of the auxin transport inhibitor 1-N-naphthylphthalamic acid (NPA).
To study proliferation due to infection of primary B-cell by GFP-EBV on B-cell, we monitored expression of the intracellular proliferation marker Ki-67 [34] by Flow Cytometry.
As for BMP signaling, we monitored expression of the Smad1-target Sizzled on the ventral side of the embryo.
We monitored expression of the reporter allele specifically using primers spanning Gata6 exon1 and the neomycin reporter gene (supplementary material Fig. S1).
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