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However, paired-end protocols virtually double the read per template and facilitate some applications that were originally out of the reach of the technology. ilmn). 4 Life Technologies SOLiD ABI (now part of Life Technologies) has commercialized the SOLiD (Support Oligonucleotide Ligation Detection) platform. This platform is based on Sequencing by Ligation (SbL) chemistry. SbL is a cyclic method but differs fundamentally from other cyclic NGS chemistries in its use of DNA ligase instead of polymerase, and two-baseencoded probes instead of individual bases as units.
After analyzing the results in five categories individually, it was determined that three of the categories, amplicon yield, amplicon quality, and percent HQ, were most affected by the variables being investigated. Therefore the influence of these categories on one another was more thoroughly studied to discern the most optimal percent 7-deaza-dGTP mixture. Figure 5A(I to V) shows scatter plots of the percent HQ and amplicon yield for all variables being tested. The shaded portion of the plot highlights dNTP mixtures that reached a threshold of at least 50% relative amplicon yield and 50% high quality bases called.
28 DNA Sequencing – Methods and Applications In summary, these studies have investigated both the percent of 7-deaza-dGTP substitution and the influence of standard and CleanAmp™ versions of the nucleotide mix on PCR and Sanger sequencing performance. When different metrics such as amplicon yield, amplicon quality, HQ percentage and sequencing chromatogram quality are considered, there are notable instances where reactions employing CleanAmp™ dNTPs have either comparable performance or statistically significant improvements in performance.