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blue dna

FAQ

General

The main GSL facility is located on the second floor (room 2518) of Thomas Hall on the North Campus of NC State University. The GSL Plant Genotyping Facility is located on the second floor (rooms 2364 and 2368) of the Plant Sciences Building on Centennial Campus. For maps and contact information, visit Contact Us.

The GSL uses an online Lab Submission system for projects. For more information on access, as well as a user’s guide, please see our How To Submit Projects page.

Our hours for sample drop-off are 9 AM until 4 PM Monday through Friday. We ask that if you want to stop by during lunch (12-1 PM) that you call ahead to confirm your plans (919-513-3882).

Of course! We encourage you to discuss your sequencing project needs with us. Please email or call Andy (919-513-0738; dabaltze@ncsu.edu) or Kelly (919-515-2012; kafridey@ncsu.edu) to set-up a time.

Yes. We have many customers from all over the country and around the world. Please call or email Andy (919-513-0738; dabaltze@ncsu.edu) for questions about billing concerns.

All academic or U.S. federally-funded parties are currently charged the same use rates. Other parties or businesses, please Contact Us to discuss pricing.

We are strictly a wet lab and unfortunately do not offer bioinformatic services. Our endpoint deliverables are demultiplexed .fastq files (Illumina) or .bam files (PacBio). Please contact the NC State Bioinformatics Consulting and Service Core (genomicsci-support@ncsu.edu) to schedule a consultation to discuss your research needs after receiving your data. 

We do offer a draft contig assembly for our Phage Sequencing Service using CLC Genomics Workbench. 

Sanger DNA Sequencing

For Gold service, we typically require the user to add specific primers to their DNA template, for each reaction. For Platinum service, as we perform Exosap cleanup of the PCR template, we therefore request users provide their primer in a separate tube which will be added following the ExoSap step. More information about these specifications can be found on our Sanger DNA submission guidelines page.

The GSL does maintain stocks of common vector-based sequencing primers, in which users can request to be added (either Gold or Platinum service). These primers are M13F, M13R, T7, or Sp6.

M13F(- 41): 5′ – CGC CAG GGT TTT CCC AGT CAC GAC – 3′
M13R(- 48): 5′ – AGC GGA TAA CAA TTT CAC ACA GG – 3′
T7 Promoter: 5′ – TAA TAC GAC TCA CTA TAG GG – 3′
SP6 Promoter: 5′ – TAC GAT TTA GGT GAC ACT ATA G – 3′

For Sanger DNA submissions, for 8 or less samples, users may submit their samples in clearly labeled 1.5 mL microcentrifuge tubes or PCR strip tubes. For more than 8 samples, please submit your samples in a 96 well plate (even if requesting < 96 reactions) or use 8-strip pcr tubes/caps. Users submitting samples by FedEx/UPS delivery should ensure the samples are adequately sealed and protected during shipping.

**Please note only Gold and Bronze Sanger Sequencing services are available for next day service** 

For same day data delivery, we must have your Lab Submission Order no later than 9:30 AM on the day of service, and samples must arrive at the GSL by 10:30 AM that same day. Orders/samples received after these times will be delivered the following business day. Users requesting our on campus courier service will need to have their samples in the courier dropbox no later than 10:00 AM.

For Gold and Bronze services, if you meet the time criteria for Order submission and sample drop off (see above FAQ), you should have results sent by email by around 5 PM the same day. If you were not able to meet this time criteria, or are submitting samples for Platinum sequencing, you should get results by 5 PM the next business day.

Note: The GSL will make every effort to adhere to this regimen, however there are times when data delivery may be delayed due to factors beyond our control (instrumentation issues, supply backorders, or a higher than expected submission volume). We cannot fully guarantee same-day delivery for your samples.

Next Generation Sequencing

Please see our NGS Library Prep page for more information on the types of NGS library preparation services we offer.

Although it depends on the specific type of NGS library prep you are interested in, a good starting point is 1-2 μg of HMW DNA or RNA in a volume of 40-50 μl (20-50 ng/μl). If you are unable to prepare this amount, or need more information on specific library prep requirements, please contact the GSL team.

We do accept user prepared libraries for sequencing, under the following criteria.

For Illumina libraries, we require a final pooled library at 10 nM (we do not offer pooling services). For each lane or flow cell requested, please provide a volume of 30 μL. If your library is significantly lower than 10nM, please contact us to see if it can be sequenced. Note: if requesting a full NovaSeq S4 flow cell, please submit a volume of 120 μL (4 XP lanes per flow cell).

For PacBio libraries, please provide a final library pool at 20 ng/μL with at least a 10 μL volume.

** Disclaimer 1** – the GSL adds a mandatory QC charge of $32.00, which is necessary for proper loading. While we will report any issues we see with the library QC spectra to customers before sequencing, the GSL cannot guarantee results for libraries we do not make, and will need to charge regardless of the outcome.

** Disclaimer 2** – for single XP lane options (Illumina NovaSeq 6000), customer libraries must allow standard Illumina index and read cycling settings, with no custom primer addition, as these impact other lanes on the flow cell. Whole flow cell selections do not require this. Please Contact Us for more information.

We currently house two Illumina MiSeq sequencers, one NextSeq 500 sequencer, one NovaSeq 6000 sequencer, and one PacBio Sequel in our facilities. Please see our NGS Platforms page for more information on these types of NGS sequencers.

We recommend shipping DNA overnight with freezer packs. For RNA we recommend shipping on dry ice, with the total RNA in nuclease-free water (50 μl). If the sample is mRNA, please contact the GSL to determine the appropriate volume.

Please see our Contact Us page for facility shipping information.

DNA & RNA Extractions

The GSL offers high-throughput DNA extraction services using either the LGC Oktopure or Thermo Kingfisher robotics platforms. For more information, please visit our HT DNA isolation page, as well as our DNA extraction guidelines.

The GSL does offer RNA extractions under certain conditions. Please see our RNA extraction guidelines for more information on this service.

Bioanalyzer, Tapestation & BluePippin

The GSL offers a variety of à la carte QC services for DNA and RNA. Please visit our DNA/RNA Sample QC page for more information.

No. For NGS project submissions, Bioanalyzer & Tapestation QC components are included directly in the NGS form. Users should only fill out a Bioanalyzer/Tapestation request form if they just want QC services (for example, if you are testing different isolation protocols or making your own NGS libraries). The GSL only accepts small, disposable sample aliquots for this service.

For the RNA Pico chip, please bring your samples at a concentration of 1-5 ng/μL. For the RNA Nano chip, samples may be at 50-200 ng/μL.

We offer five different Tapestation assays. For HMW genomic DNA (100 bp to 60 kb) please bring your samples at a concentration of 10- 100 ng/uL. If you are interested in fragments between 100 and 1000 bp, samples should be at 0.1 – 50 ng/uL for the D1000 assay or 10-1000 pg/uL for the High Sensitivity D1000.  If you are interested in fragments between 100 and 5000 bp, samples should be at 0.1 – 50 ng/uL for the D5000 assay or 10-1000 pg/uL for the High Sensitivity D5000. 

Please bring 3-5 μl of each sample, at the appropriate concentration, in 1.7 mL tubes, labeled clearly. For picogreen or HT services, please submit your samples in a 96w plate, arranged by column (not rows).

BluePippin samples should be submitted in exactly 30 μl of EB, TE, or water. For more information about this service, please visit our BluePippin page.

Other Questions

While many of our instrumentation can be used only by GSL staff, we do operate and maintain shared user equipment at both our main GSL facility and the Plant Genotyping facility that is available for use after training. Please see our Shared User Equipment section for more information.

Training requests and use scheduling are made through the GSL Lab Submission system. For a user’s guide to using this system, please see our How to Submit Projects page.

No. We offer qPCR instrumentation use as shared equipment, and our customers set up their own assay plates and run them. Training requests and use scheduling are made through the GSL Lab Submission system.

Currently we cannot offer direct in person training due to our work volume. We can provide customers with a review of relevant protocols, and further Q&A. Additional opportunities for NGS library prep training, as well as other applications, may be available periodically through the NC State University Biotechnology program.