Sample Requirements

Sample Requirements | Isolation | Quality Control | Submitting Samples

GSL pricing is based on the assumption that submitted samples can go straight into reactions, requiring little to no manipulation of samples by a technician. Therefore, any samples submitted that do not meet the requirements below will be subjected to a $25 surcharge per sample. If you have questions about our requirements, please contact us before isolating or preparing your samples for submission.

Heparin is a powerful PCR inhibitor and co-purifies with DNA. Therefore, DNA extracted from blood stored in Heparin-coated tubes should not be submitted.

  1. Samples for isolation: The GSL accepts a variety of material for DNA or RNA isolation, including blood, saliva, cells, tissue, buccal swabs, and fecal matter. Tubes containing material for isolation must be clearly labeled with the GSL project and sample number ("1234-SL-0001"). Please discuss your isolation requirements with us before submission of material so we can advise you on the best isolation procedures to be used for the goals of the experiment.
    • For standard genomics applications such as exome sequencing, non-phased whole genomic sequencing, or RNA-seq, blood should be collected in an EDTA (lavender top) tube. Heparin-containing tubes should not be used. For RNA extraction, it is advantageous to submit tissue or blood in Trizol or RNAlater.
    • For saliva collections, the GSL prefers Oragene Dx (for Diagnostics).
    • For buccal swab submissions, we recommend the mawi iSWAB-DNA Collection Kit.
    • Please note that the long-range phased DNA sequencing platform from 10X Genomics requires high molecular weight (HMW) DNA that cannot be obtained from standard isolation techniques. Specialized protocols must be used to obtain HMW DNA and a CPT blood collection tube with sodium citrate must be used for blood submissions. See our 10X Genomics platform services for more information.

  2. Buffer: Samples may be suspended in a wide variety of buffers, including 10mM Tris, dH2O, or Qiagen EB. The most critical requirement is that buffers contain no more than 0.1mM EDTA. For RNA extraction, it is advantageous to submit tissue or blood in Trizol or RNAlater. Certain applications have specific submission requirements, such as 10X Genomics platforms requiring High Molecular Weight DNA. If you have questions about submission requirements for a specific platform, please contact us before submission online.

  3. Labware: All samples should be submitted in either 1.5mL centrifuge tubes (9 samples or less) or polypropylene PCR plates (10 samples or more). If submitting in tubes, write the sample number listed on your order sheet on the top of each tube. Other identifiers may be written on the side. Plates should have the project number and sample number range clearly marked, and should be sealed firmly with a foil plate seal capable of withstanding -80 C temperatures, such as the Corning Micoplate Aluminum Sealing Tape Product 6569). We prefer this semi-skirted plate # AB-2400 in clear. Any plate submitted must be CLEAR and semi-skirted or full-skirted. No deep well, round-bottom, or flat-bottom plates!

  4. Protecting Your Samples: We recommend the following products to seal your microplates for shipment:

    BioRad Film Sealing Roller for PCR Plates
    Adhesive Seal Applicator

    For shipping on dry ice, we recommend placing your plates or tubes in a freezer box (with or without a divider), for example:
    Freezer Box with Dividers

  5. Concentration & Volume: Required amounts for various applications can be found in the table below. To avoid handling fees of $25/sample, the minimum required amount must be sent at the concentration requested, in a minimum volume of 15µl. Prepared libraries may be sent at 15 nM if concentrations were determined by fluorometric means (picogreen/Qubit) and QC metrics are sent (bioanalysis PDF). The most frequent sample-related problem we encounter is low concentration, particularly with dsDNA. If samples are precious, please contact us prior to submitting.

All samples undergo a thorough quality control before being submitted for each application. The inputs listed should remain after the completion of QC. For RNA samples please include enough volume to allow for 2-4 uL to be used in QC. For DNA please provide enough volume to allow for ~2-8 uL to be pulled. The volume of sample needed varies with concentration.

RNA Sequencing PolyA Ribosomal RNA Reduction Small RNA Low Input
Warrantied input 500 ng in 50 µl 500 ng in 28 µl 1 µg in 6 µl 1 ng -100 ng in 5 µl
Lowest input attempted 100 ng 100 ng 100 ng in 6 µl 500 pg/high RIN
Recommended concentration > 20 ng/µl > 20 ng/µl > 167 ng/µl > 1 ng/µl in 5 µl
Recommended integrity value (RIN) > 7 > 7 ideal; < 7 acceptable > 7 > 7
DNA Sequencing Standard WGS/HiSeq X WGS Chromium by 10X Exome Capture Mate Pair WGBS
Warrantied input 1000 ng 500 ng in 20 µl 1500 ng 1µg (gel free); 4 µg (size selected) 200ng
Lowest input attempted 500 ng 1 ng 500 ng 800 ng 200 ng
Recommended concentration 100 ng/µl 500 ng in 20µl 100 ng/µl 100 ng/µl > 10 ng/µl
Other Services Library Submission Illumina MethylationEPIC BeadChip Illumina Omni BeadChip Illumina HT-12 v4 BeadChip Affymetrix PRIMEVIEW
Warrantied input > 15 nM in 15µl 1 µg in 45 µL 250-300 ng in 6 µL > 500 ng in 11 µL 100 ng in 3 ul
Lowest input attempted 1.0 nM 500 ng in 45 µL 200 ng in 6 µl 500 ng 50 ng
Recommended Concentration > 15 nM > 25 ng/µl > 50 ng/µl > 50 ng/µl > 33.3 ng/µl
Recommended integrity Robust kapa qPCR with smooth profile Intact Intact RIN >7 RIN >7
Considerations Minimum batch is 32 Minimum batch is 48 Minimum batch is 12
ChIP Sequencing
Input recommendations 100 ng in 44 µl
Lowest input attempted 0ng/µl
Size recommendations prior to IP 250-300bp, maximum of 600bp

*GSL ChIP-seq policy: ChIP-seq experiments are by nature extremely complex and contain a large number of variables, including DNA sonication sizes, cross-linking protocols, immunoprecipitation efficiency, post-IP purification, and limited quantities of DNA produced. If we are not able to quality control the submitted ChIP DNA, it is impossible to predict the performance of an experiment, both from a technical standpoint (i.e. will a high-quality sequencing library be produced) and from an analysis standpoint (i.e. will the results show expected, high quality data). For any samples that do not meet input specifications, or are not able to be quality controlled due to limited DNA amounts or volume, we will not warranty any technical aspects or analysis performance. For ALL samples submitted for ChIP-seq, we make no warranties for analysis performance, meaning that we do not guarantee the results will be what are expected or are valuable because we have no control over the highly complex nature of the ChIP portion of a ChIP-seq experiment. We can only work with the DNA we receive, and since we do not know the true nature of what is received, we cannot predict the outcome. For all but the most experienced labs who have a long track record of success with ChIP technologies, we recommend using a full-service provider such as Active Motif.