SUBMISSION IN 3 EASY STEPS

MEASURE

Plasmid: Sample concentration

  • Plasmid DNA samples should be submitted at 5.6 ng/ul in > 20 ul volume. Preferably, DNA should be submitted in nuclease-free water. If DNA is in an elution buffer, such as 10 mM Tris-Cl, pH 8.5 (Buffer EB), please dilute the sample in nuclease-free water to the required parameters.
  • Measure the DNA concentration using Qubit or similar fluorometric methods and normalise your samples to 5.6 ng/ul in > 20 ul volume. Try to be as close to these parameters as possible. Samples with too little OR too much DNA may result in sequencing failure.
  • The QuantIT kit uses the same chemistry but can be adapted for plate reader based measurement of up to 384 samples at a time
  • IMPORTANT: Nanodrop and other spectrophotometric instruments are NOT accurate enough to quantify DNA for nanopore sequencing. It is imperative to submit your sample at the required concentration and volume as it will otherwise lead to an unsuccessful sequencing run.

Plasmid: Sample purity

  • Determine DNA purity on a Nanodrop spectrophotometer (or similar spectrophotometer). DNA parameters should be within the following ranges:
  • Absorbance ratio 260/280 (indicator for protein contamination): ≥ 1.8.
  • Absorbance ratio 260/230 (Indicator for organic compound or chaotropic agent contamination): 2.0 - 2.2.
  • Low quality samples outside of the parameters above may result in sequencing failure.
  • For more information on 260/280 and 260/230 ratio interpretation see awhitepaper from Thermo Fisher Scientific

Amplicon: Sample concentration & purity

  • Run your PCR reaction on an electrophoresis gel to confirm a strong band at your expected amplicon size.
  • No dilution and no purification is necessary. Simply prepare your samples for shipping as stated in the “Pack” step below.

PACK

Please submit your samples only in PCR strip tubes or 96-well PCR plates (or single PCR tubes if submitting a single sample). For 96-well PCR plates, add your samples from top to bottom and left to right (i.e.: 1st sample in well A1, 8th sample in well H1, 9th sample in well A2, and so on). Other types of tubes will not be accepted and might get discarded.

Register samples

  • Create a new order. Once you have registered your order a 3-letter order code will be generated for you.

Label samples

Single PCR tube

  • Label the tube with the 3-letter order code.
    • a single PCR tube with ABC written in blue marker on the side

PCR strip tubes

  • Label the first tube of each strip with the 3-letter order code.
  • Label the second tube of each strip with the strip number. (I.e.: Label the strip containing your first 8 samples with “1” and repeat for remaining strips in the order you registered your samples).
    • a row of 8 PCR tubes with ABC written in blue marker on the side of the first tube and a '1' on the second tube

96-well PCR plate

  • First seal your plate with a heat seal or strip caps (See step 3 below).
  • Add your samples to the plate in columns not rows, as specified in your order submission.
  • Label the plate with the location of your first and last sample.
  • Lastly, label with the 3-letter order code and plate number (If submitting multiple plates per order)
    • a full 96 PCR well plate with a foil on top. On the foil is written with a blue marker 'ABC'. The first well in the top has written '1' and the last well that contains a sample '48'
  • Don’t forget to label your samples, as we otherwise have no way to determine the samples are yours!

Wrap samples

Single PCR tube or strips

  • Tightly close lids.
  • Wrap tubes in light layer of parafilm.
  • Place in protective vessel (e.g.: 50 ml Falcon tubes).
    • a single PCR tube wrapped a strip of PCR tubes wrapped
    a singe wrapped PCR tube in a Falcon tube a strip of PCR tubes wrapped in a Falcon tube

96-well PCR plate

  • It is crucial to ensure the plates are sealed extremely well.
  • Seal plates with a heat seal or strip caps. Adhesive seals are NOT recommended.
  • Use of a roller can help to ensure a tight seal.
  • Wrap the plate in packaging material (e.g.: bubble wrap) to prevent cracking during transit.
SHIP

Sample shipping

  • Together with your samples include a sheet of paper containing your order label, name, and sample number.
  • Dry ice or cold packs are not necessary. Room temperature shipping is absolutely fine.
  • Place your samples in one of our dropboxes or ship samples via express courier to our address shown below. If you are sending your samples directly to us, they need to arrive at our laboratories before Tuesday 13:00 or Friday 9:00 to be included in the days sequencing run. Ship samples to:

Unveil Bio ApS

Fruebjergvej 3

2100 Copenhagen

Denmark

  • That’s it! You can check the status of your order on our website. You will receive an email notification when your analysed data is ready for download on our website