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Noteably, MRE11 was detected in the DSwt, but not in any of the control templates lacking telomere repeats.
To optimise the Taqman PCR reactions and normalise the data, we generated a standard curve using the control templates.
Each carrier's data were then fit by the control templates and the resulting values for the a-scale and t-scale were plotted for each hexagon.
The control templates were two BACs spanning the FRG1 region and the D4Z4 array (RP1-226K22 anD4Z4-containingning BAC isolated from a genomic library screening) (B. Bodega, unpublished results).
BAC clones (20 μg) containing XPB/ Eccr3 (MRC Geneservice, clone 344-C18), Pitx2 (CHORI, clone RP24-215O15), Enpep (CHORI, clone RP24-172B1) and 3 G3 tested region (CHORI, clone RP23-356C23) were treated in parallel, to generate the control templates.
Comparison of these melt profiles to those of the control templates (Fig. 2a) indicated that these individuals carried 3 copies of the α-tryptase gene, suggesting that these individuals carried an allele involving duplication of the α-tryptase gene in addition to a single-copy allele.
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Moreover, A ij,kl and B ij,kl are the parameters forming the feedback template A and the control template B, respectively, whereas k l ∈ N r ̄ is a grid point in the neighborhood within the radius r ̄ of the cell ij[20].
The sequencing electrophoregram of the control template with the sequencing primer Gre is shown in Figure 4.
In standing start experiments with control template, both hpol κ (a 10 1 molar ratio of polymerase to primer template duplex) and hpol η (a 4 1 ratio of polymerase to primer template) completely extended the primer opposite the control template to form 18-mer products (X = dT, Figure 3).
Although 50 nM or higher concentration of pol IV readily extended the 14-mer primer to a full-length 23-mer product on the control template, it was strongly blocked by the cyclopurine lesions.
Consequently, the catalytic efficiency values corresponding to the control template were normalized to 1. Since each polymerase had different insertion efficiency values for a specific nucleotide and adducted template, the polymerase with the lowest insertion efficiency value was calculated and normalized to the value of 1.
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