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The majority of existing plant transformation vectors are quite large (typically 10 kb or more) and therefore may not amplify well using PCR.
Because the polymerase extension time is fixed in the library amplification step, longer fragments may not amplify in the initial PCR stages, but all short fragments will.
The main disadvantage of PCR is that certain fragments (particularly the smaller ones) may be selectively amplified, and that it may not amplify large (> 4 kb) fragments.
This technique is sensitive but provides a lower-bound estimate for activity levels because long fragments may not amplify due to PCR bias.
Intron-exon junctions at 5' or 3' UTR may not amplify well using standard PCR conditions that were likely employed by Otto et al. [ 17].
The results from this mouse study indicate that the immune-selective action of dipyridamole may not amplify the osteoporotic effect and fracture risk associated with very-low-dose prednisolone [ 48] in the clinical setting with chronic dosing.
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However, their predictions are not always reliable as elements with the same name can vary at the level of their DNA sequences in different GMOs, and a set of primers and probes designed for detecting a specific element may or may not efficiently amplify a slightly different version of the same element.
In other words, because of PCR bias, some species may not be amplified and sequenced while others may be amplified and sequenced in excess.
When the gene transcripts are amplified in a multiplex way, certain low-copy-number sequences may not be amplified to the detectable amounts when the reaction reaches a saturation point.
The production she starred in may not have amplified the soloists, as this one did, but it is doubtful whether you could have heard the children's voices without the enhancement.
This result suggests that positive GRE-mediated glucocorticoid adverse effects, including hyperglycemia, may not be amplified by simultaneous dipyridamole treatment.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com