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For the correct timing of the fixation, embryos were continuously scored for each cleavage.
For each cleavage site (site A), the distance to each of the nearby cleavage sites (site B) is recorded.
All-against-all protein product identity scores were produced using the Belvu programme [ 68], and reformatted into symmetrical square arrays of sequence identity percentage values (one for each cleavage product) represented as greyscale bitmaps in Figures 2B and 2C.
For each cleavage the residues P4 P4′ were highlighted to indicate whether the residue in each sequence had been observed in any substrate at that position for the peptidase in question.
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The calculated cleavage energy for each possible cleavage site shows that Ti Si and Ti Al are the weakest layers, while the Ti C layer is the strongest one.
Our data support this mechanism, as the kinetic values for each short cleavage sequence do not correlate with the order of the cleavage.
Associated formulas with rearranged hydrogens are shown for each labeled cleavage Table 2 Summary of sphingolipid classes and MS/MS spectra developed in this study.
The residues from P4 P4′ were collected for each substrate cleavage.
Molecular weights for each FeBABE cleavage fragment were determined by measuring the migration distance, obtaining the log10 value from the straight line fit of the molecular ruler, and taking the inverse log (10 n ).
Nonetheless, the levels of targeting observed in this study have been obtained with a minimal system that relies on simple base pairing to a guide RNA, in contrast to the ZFN and TALEN proteins, which require a new protein to be engineered for each new cleavage site.
Deep sequencing of the resulting fragments generates, for each RNase, a cleavage profile along the mRNA sequence.
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