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Consistent amplification was achieved with only one pair of primers for which the PCR conditions were further optimised.
From this dataset, a set of primer pairs (15) was validated by polymerase chain reaction with 86% consistent amplification, confirming the accuracy of the in silico prediction.
Chromosome location 8q24.12 approximately q24.13 was the only region that showed consistent amplification in approximately 50% of the HER2- cases.
Importantly, consistent amplification patterns were observed within a given Tm.
Standard curves were run for all assays to ensure consistent amplification efficacy.
In contrast, PCR performed on 10 sorted cells led to consistent amplification of the cellular gene (Fig. 3B).
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To obtain reproducible and clear banding patterns, each amplification was repeated three times, and only bands showing consistent amplifications were scored.
Of the 88 SRAP primer pairs, 20 primer combinations that produced consistent amplifications and clear polymorphic bands were selected to amplify the rest of the samples (Table 2).
The diagnostic LTR-PCR used in this study only yielded consistent amplifications when using CNS as DNA source, since all animals were seropositive and yielded PCR-specific amplicons in CNS (Table 1).
Our ILPs displayed consistent logarithmic amplification plots, demonstrating a sustained amplification efficiency, while the random pentamers with more hybridization sites led to inconsistent plots.
Consistent Kv1.1 amplification from cDNA prepared from rodent islets required 40 cycles of amplification from template levels corresponding to 50 ng total RNA.
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