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We constructed two mutants with one or two non-polar bulky residues (I or IF) inserted after A129 (A129InsI and A129insIF) to lengthen the loop between H4 and H5.
In contrast, no increase in these parameters was observed for yar mutants, with one exception.
We generated three mutants with one or both acidic stretches deleted, as well as three swap mutants within the stretch and confirmed the proposition.
To investigate whether zebrafish mutants with one wild-type ptena allele (ptena +/−ptenb−/−) develop tumors during their life span, we monitored around 300 ptena +/−ptenb −/− mutant fish.
Interestingly, Cas9 nickase greatly reduces the ability to generate mutants with one sgRNA, suggesting that the application of Cas9 nickase and sgRNA pairs can almost avoid off-target effects when generating indel mutants.
In this respect, analysis of double mutants (with one mutation being the rat1-1 mayation) may be instrumental in finding additional regulators of TERRA stability and, furthermore, of TERRA transcriptional activation.
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The sequenced fragments or markers were completely homozygous in Ugni Blanc and Ugni Blanc mutant, with one marker analyzed each 23 kb in average.
Since siRNAs can have off-target effects, it is important for functional assays to make a specific mutant with one or more base mismatch within the target recognition site as a control [ 21].
We found that the wild-type MIR-199A2 promoter showed elevated transcription during DCA stimulation, while mutant promoters with one or two mutant AP-1 binding sites had significantly weaker activity.
Both mutants with only one exposed cysteine residue (63A/224C and 63C/224A) reacted with only one 5-IATR molecule per protein (Supplementary Table S1 at http://www.BiochemJ.org/bj/440/bj4400043add.htm).htm
In this assay, a mutant DNA with one base mismatch (C→G mutation) is first hybridised to a complementary DNA modified magnetic NP.
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