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Therefore sequences can rapidly be sorted into their original template source using the tags (Figure 1).
Then, depending on template source, 40 or 45 cycles of 95°C for 30 seconds, 55°C for 1 min and 72°C for 1 min followed and the reaction ended in a final extension of 10 min at 72°C.
In this paper we have overcome this problem, presenting a method where initial PCR primers are 5'-tagged with short nucleotide sequences (tags) in such a way that a unique tagged primer combination can be applied to each specific DNA template source.
Mostly pooled cDNA samples from various tissues were used as a template source.
However, they used their own GIS flcDNA library as the template source.
Also provided are the target gene name/annotation, template source for PCR and purpose of amplification.
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Free from expensive template sources and complex manipulation, this work might shed some light on the synthesis of low-cost and high-performance materials for the next "beyond lithium" battery generation.
Profunc [33] mitigates these problems by using multiple template sources, including enzyme active sites gathered from the Catalytic Site Atlas [30], ligand and DNA binding sites [34], and automatically generated triads of amino acids called 'reverse templates' [33].
Since sample variations or differences in sample preparations may influence any observed difference in enzyme efficiency [27], we used a well tested primer pair and viral RNA standard, the Armored HIV RNA [28], [29], thereby eliminating potential variation due to template sources.
This approach also eliminates template source-based variation in PCR amplification, a significant cause of assay nonlinearity.
Three additional species were later added to the project when DNA and RNA template sources became available, and a full list of the 16 targeted species and their putative evolutionary relationships are depicted in Figure 1.
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