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As the coding sequences of two subfamilies are very different we were not able to design a common primer sets for the family.
In the case of SOX2 and OCT4 genes, due to their high sequence conservation between human and mink in the coding region, we were not able to design human specific primers.
Although genes C13ORF18 and NKX6-1 were part of combination QMSP assays of interest [10] we were not able to design corresponding pyrosequencing assays due to high CpG density.
For a number of microsatellites detected computationally, we were not able to design primers because they either lacked suitable flanking sequences or the total sequence length was too short (< 120 bp).
Likely because of the repetitive sequences in the flanking region of rs7871490, we were not able to design a pyrosequencing assay, which precluded a direct comparison between SNaPshot and pyrosequencing.
Previously, we have explored the problem of aligning two sequences simultaneously to a profile HMM, but we were not able to design a simple generative model for this purpose [ 16].
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This challenge was often described as a reason why program planners are not able to design comprehensive services, and also as a contributor to overlapping efforts and inefficient use of resources.
Because of this limitation, we were not able to optimally design the primers based on a balanced melting temperature (Tm).
If you were not able to find an appropriate design from the sample provided by the supplier, do not worry because you can use a touch of creativity in coming up with your design.
As we included exclusively BP with objectively assessed neurocognitive deficits, recruitment was difficult and subsequently we had a small sample size and were not able to implement a randomized group design.
First of all, despite our best efforts, we were not able to build a completely satisfactory design.
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