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For ChIP PCR, primers were designed using Primer3 software according to the following criteria: expected amplified fragment size between 150 and 300 bp, primer size between 18 and 25 bases, and primer melting temperature (Tm) between 55 and 60°C.
RT-PCR showed the expected amplified product of 1347 bp for TCII-OLEO, 1240 bp for OLEO-TCII, 551 bp for TCII, and 275 bp for OLEO (Fig. 1A).
The expected amplified fragment for IGFBP-2 was 162 bp.
The expected amplified fragment for GAPDH was 259 bp.
The Hygr transformants were PCR screened with gus- gfp primers which resulted in an expected amplified product of 2.5 kb.
The expected amplified sequences encompass exon 3 and exon 4 and corresponded to the C-terminal region of the extracellular domain of these receptors.
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As expected, some amplified genes (MEMO1, IGFP6, TBK1, MIB1, RIOK3), determined by the DNA chip array demonstrated high level of RNA overexpression as well.
As expected, the amplified products were assigned to rat fut8 transcripts (NW_047761.1, NW_047771.1) excepted for the 450 bp band, which was not specific.
In particular, strong evidence for deletions is provided from genomic regions with too-low coverage and average mapping distance greater than expected, while amplified regions entail extremely high coverage and average mapping distance less than expected [ 3].
But one half expects an amplified John Williams soundtrack to punctuate the overblown run-on rhetoric.
The sequences of three pairs of specific primers were used to identify genes Bin A, Bin B that encode binary toxins (41.9 and 51.4 kDa) and gene Mtx1 that encode mosquitocidal toxin, 100 kDa; the expected size of amplified products are shown in Table 1.
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