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Despite using high-fidelity long read DNA polymerase and specific PCR conditions, we were not able to consistently amplify the first two exons, probably because of the very high CG content of this zone.
Samples that did not easily or consistently amplify were re-extracted using a commercially available kit (Qiagen) and this DNA source was used instead.
The RT enzymes varied considerably in their ability to reverse transcribe Armored HIV RNA spiked with a typical FFPE tissue extract, showing seven of the enzymes to consistently amplify 1,000 copies of Armored RNA tested (Figure S2).
While the majority of the total eleven enzymes failed to consistently amplify across the range tested (Table 2), the five which performed well at the lowest template end tested (Figure 1), are more suitable for studies aiming at detecting low amounts of RNA templates.
FCA649 had the highest error rate and was the most difficult to consistently amplify.
The results presented in Table 3 show that of the 11 loci tested, only eight consistently amplify across species boundaries.
Similar(45)
All the viral RNA tested in this study could be consistently amplified by the assays.
In spite of initially using primers designed for Gammaproteobacteria, we consistently amplified sequences from a different bacterial phylum: Bacteroidetes.
Respiratory rate increased in both genetic groups from six to eight weeks of age, but the increase was more pronounced in Botucatu rabbits than in crossbreds, and the difference between groups was consistently amplified over weeks.
The British tenor's voice, though sometimes hard to hear in the lower register, has retained its salty-chocolate tang; in a truly radical production it would have been consistently amplified, making this newly larger-than-life protagonist a true competitor with the orchestra.
Primers Cct1 and Cct2 consistently amplified a single product of 360 bp from DNA prepared from 20 isolates covering the principle range of the disease from Central and Northern Europe, but not from other Neonectria, 'Nectria' or a range of species commonly encountered in forest ecosystems, as well as P. abies or P. radiata DNA.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com