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Mass spectrometric analysis of biotinylated ThioFab (Fab with engineered cysteine) variants confirmed that conjugation occurred only at the engineered cysteine thiols of either light or heavy chains.
Moreover, the screening of control region polymorphisms that are diagnostic for M23 against a larger global panel of mtDNA control region variants confirmed that the M23 control region motif is indeed rare, as only four individuals shared the 13 control region diagnostic mutations for M23 (Additional file 1).
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The potential reproduces both the measured 14CH4 isotherm for PCN-14 and the site preference for adsorption at OMS. Extension to 5 metal-substituted variants confirms that OMS composition can dramatically alter uptake, with Ni- and Ca-based compounds predicted to exceed the performance of Cu-PCN-14 largely.
The intracellular eukaryotic expression of αRep variants confirms that they can be stably expressed in mammalian cells.
Fitting the t lag (pHdrop hemifusion ) distributions with a gamma probability density yields unchanged values of N (between 3 and 6) for all the variants, confirming that what changes with pH is not the number of rate-limiting rearrangements leading to hemifusion but only their rate.
The increase in MVD values from luminal to mural variant confirms that the mural variant is more belligerent and prone to recurrence, while the intraluminal and luminal variants are thus amenable to treatment by enucleation and curettage.
Genomic DNA sequencing was also performed on heterozygous sequence variants to confirm that identical sequence alterations were present in genomic DNA.
While a knockout model cannot confirm that the results are not due to a variant within the HFE gene in LD with the H63D/C282Y variants, it does confirm that LD with a variant outside the HFE gene is not driving the results.
Our intuition is that that the variants confirmed in both samples are more likely to be genuine germline polymorphisms.
Furthermore, analysis of the sequences of the resulting splicing variants confirmed our previous observation [ 2] that most of the investigated AS events resulted in introduction of at least one in-frame PTC.
An excess of homozygotes can also be observed when there is population substructure; however, as a control, we show that the variants confirmed previously (2, 7) had fewer deviations from HWE than our unfiltered, raw variants (Supplementary Material, Fig. S3), which is why we conservatively applied the Hardy Weinberg filter.
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