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The throughput of this assay is high and suitable for HTS screening.
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Additionally, the high throughput nature of this assay lends itself to the determination of kinetic parameters for each target site, allowing us to determine the kcat/ K1/2 of either protein for a given site.
In order to increase the throughput of the assay, two multiplex pyrosequencing assays were concomitantly developed using three and two pyrosequencing primers (Table 2) to generate corresponding sequences for blaTEM and blaSHV amplicons, respectively.
In this study, we evaluated the feasibility of automating the serum RNA extraction procedure in order to increase the throughput of the assay.
Additionally, the cumbersome manual loading process significantly limits the overall throughput of the assay (∼5 min loading out of 10 min total assay).
The addition of an immunoaffinity enrichment step to the multiplex detection scheme allowed us to circumvent the time-consuming electrophoresis step and improved the throughput of the assay.
Yet, MS methods have the drawback that they often require tedious protocols for sample preparation that decrease the throughput of the assay and significantly increase the cost.
A second drawback was our reliance on manual spotting for the replication of bacterial colonies, which severely limited the throughput of the assay and increased the likelihood of human error.
We next used a microfluidic device to produce single- to subnanoliter DEX droplets to increase the throughput of the assay and demonstrate the potential for automated, chip-based analysis.
MRM-based quantitation can also be multiplexed to analyze and quantify hundreds of proteins per run, increasing the throughput of this type of assay and making it fast enough for clinical applications.
At present, SNPs are used most frequently because of coverage of a large fraction of genome, throughput of assay, quality assurance and cost-effectiveness.
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