The threshold cycle ( C T) values of these samples ranged from 23.91 to 35.11. A total of 31 DENV-positive serum samples belonging to 4 serotypes and 9 genotypes of DENV were involved in the validation of the primer panels. In this study, we report the improvement of the existing WGS process for DENV by optimizing the primer design procedure, designing serotype-specific primer panels and reducing the sizes of amplicons. The development of next-generation sequencing (NGS) allowed its application to viral whole-genome sequencing (WGS). Historically, molecular epidemiological studies of DENV depended on E gene sequencing. The GSL does not perform assays on this equipment as a direct service.Dengue virus (DENV) is the most globally prevalent member of the genus Flavivirus in the family Flaviviridae, which can be classified into four serotypes. Please contact us before purchasing your own supplies for qPCR to ensure you are using the correct type.ģ This is shared research equipment and available only to local users. Users are charged in increments of 0.25 hours, depending on total use time.Ģ When running the CFX384 qPCR machine, the user must use BioRad optical seals and qPCR plates. Miscellaneous Services & Supplies Serviceġ Usage fee applies to the Covaris Ultrasonicator, Qiagen TissueLyzer, Synergy H1 Hybrid Plate Reader, and GenoGrinder 2010 instruments. We cannot guarantee customer-supplied libraries will meet this estimate.ģ Lane or block pricing reflects divided user costs, however sequencing turn-around times are subject to sufficient user volume in order to fill the flow cell.Ĥ Block sequencing is performed on NovaSeq S4 150 PE lanes, and is only available for select GSL-prepared library options, we cannot offer block pricing options for user prepared libraries, or those not suitable for sequencing at 150×2 PE, or will impact the performance of other customer libraries. Please see our Bioanalyzer & Tapestation pricing for more information.Ģ Yield estimates reflect optimum values defined by Illumina and apply to GSL prepared libraries only. Per cell (10.0 Billion reads 2) – 4 independent lanesġ For GSL library preparation, an additional charge for template quality & integrity will be added. Per cell (4.0 Billion reads 2) – 2 independent lanes NovaSeq 6000 S2 100 bp SR/50 PE flow cell Per cell (1.6 Billion reads 2) – 2 independent lanes NovaSeq 6000 S1 100 bp SR/50 PE flow cell Per cell (800M reads 2) – 2 independent lanes NovaSeq 6000 SP 100 bp SR/50 bp PE flow cell Per sample Sequencing (NovaSeq S4 150 PE) Next-Generation Sequencing Illumina Services Serviceġ0X Genomics 3′ SC RNA library prep (internal users only, BSL1 only) Gold Sequencing – Seq Rxn, Clean Up & Capillary Runīronze Sequencing – Clean Up & Capillary Run Platinum Sequencing – PCR clean up (ExoSap), Seq Rxn, Clean Up & Capillary Run Sanger Sequencing & Genotyping 96-well plate Service Bioanalyzer RNA chips run 11-12 samples, depending on the sensitivity level (pico vs nano). TapeStation High Molecular Weight DNA AnalysisīioAnalyzer RNA Analysis – Pico sensitivityīioAnalyzer RNA Analysis – Nano sensitivityġ Split pricing is available for half (1-6) and full (7-12) sets of samples. TapeStation High Sensitivity DNA Analysis TapeStation DNA Analysis (D1000/D5000 Tapes) Note: GSL DNA extraction services may not be suitable for NGS applications requiring extremely high molecular weight DNA inputs, such as PacBio or Oxford Nanopore long-read sequencing approaches. bcl file delivery available due to specific demultiplexing requirements. Due to unique cycle settings, 10X libraries require a dedicated flow cell for sequencing. Notes: 10X library prep is currently only available for internal users, utilizing cells with BSL1 (nonpathogenic) status. We will try and point you to a suitable provider if we are unable to provide services directly.Ĭurrent GSL Services & Pricing (effective April 3rd, 2023) Please contact the GSL Director BEFORE submitting ANY samples or research projects related to the SARS-COV2 pandemic or other samples of pathogenic risk to humans. The GSL is a University BSL-1 laboratory, classified as a teaching and training facility with student interactions. A standard 3% charge will be added for credit card transactions. Note: All prices below are for all researchers, including those affiliated with NCSU, UNC, Duke, and other federally-funded or non-profit organizations.
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