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This assay was applied to all Taspase1 mutants as outlined below.
It has to be mentioned that the severity of the ocular defects varies among the different Col4a1 alleles, but there is also some phenotypic variability within each mutant line similar to the features observed in our Pxdn mutants as outlined in Tables 1 and 2. In addition, mutations in Col4a1 (44– 46) and Col4a2 (47) can cause hemorrhagic stroke, but similar changes in Pxdn mutants were not found.
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To ensure that effect of the R1039A mutation is indeed measured in trans, we additionally disabled the catalytic site of the R1039A mutant by the H1061N mutation, as outlined in Figure 4a.
Samples were chosen from a resource of approximately 20000 individual M2 mutant lines (as outlined above).
Thus, future studies may wish to investigate the effect of CRY2 knockdown in mutant p53 cells, especially in light of the recent evidence suggesting differential effects of Cry mutations against the p53 mutant background, as outlined above.
The thermostabilities of HDAC8 mutants were assessed using a thermal shift assay as outlined in the Supporting Information.
As outlined in Materials and Methods, mutants MA2, MB, and MA2B were designed to fully reintroduce the Mn2+A, Mn2+B, and binuclear Mn2+A Mn2+B sites, respectively, into TbARG.
Δ vvd, Δ frq, Δ wc-1 triple mutants were identified by a PCR-based strategy as outlined above using genomic DNA isolated from pale and arrhythmic progeny arising from the cross.
As outlined above, the weak clamp mutant (Q182A) more easily allows translesion polymerases to associate with the tripartite complex.
To construct conditional knockdown mutants for ICL and Rv3671c we cloned icl1 and rv3671c as outlined in Figure 1 and integrated the expression vectors into the chromosomes of the knock out strains Mtb Δicl1 Δicl2 and Mtb Δrv3671c, respectively.
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