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Each digested sample was compared alongside with its not-digested cognate control.
The detection limits for td = 5 min in the 10 times diluted, UV digested sample were (ng L−1): Cd 7.0, Pb 1.2 and Cu 6.6, which are well below currently applied methods.
Each digested sample was analyzed separately.
Each digested sample was diluted by a factor of 3 using 0.1 % TFA.
Each digested sample was labelled by random priming using Cy5 for the trisomic samples and Cy3 for the controls.
Alternatively, we searched the chymotrypsin digested sample with low chymotrypsin specificity (cleavage carboxyterminal to L, M, W, F, and Y allowing a maximum of four missed cleavages).
The only differences were the peaks at 2θ = 20°, and 2θ = 23°, which were slightly lower and higher, respectively in the batch digested sample.
The methylation percentage was then calculated for each probe as the ratio between normalized probe peaks from the undigested sample and the corresponding HhaI digested sample.
After cooling, the digested sample was filtered and the digest was maintained to 50 ml with distilled water.
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Interestingly, the glycoform ratio of the nonPK-digested sample from the Q95H/G96S animal differed from the other genotypes, lacking the mono- and un-glycosylated PrP isoforms.
A fragment was considered to be methylated when the fragment derived from the MspI-digested sample did not correspond to that from the HpaII-digested sample.
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