High Molecular Weight DNA Extraction Guidelines

The following guidelines apply to those submitting biological samples for DNA extraction. These guidelines are appropriate for whole blood, animal tissues, animal/bacterial cells, and plant tissues where resulting high molecular weight DNA will be used for long-read applications on Oxford Nanopore and PacBio instruments.

All General Guidelines must be followed in addition to the guidelines specific to the DNA extraction.

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Biological Safety - General Guidelines

  • We are designated a BSL-2 facility. We can only accept samples that require BSL-1 or BSL-2 containment.
  • Adhere to local and federal regulations when transporting and shipping biological samples.
  • Samples requiring higher than BSL-2 containment or samples delivered to us in an unsafe manner will either be refused or disposed immediately.

Sample Delivery Vessel- General Guidelines

  • The input type and the DNA extraction requested will determine the type of storage vessel. Please see the sample-specific guidelines.

Sample Naming Conventions- General Guidelines

  • Sample names must be de-identified. Samples with Protected Health Information won’t be accepted.
  • Each sample must be identified with a simple, unique identifier. If names have been used in previous submissions, don’t repeat in subsequent submissions.
  • The following conventions are strictly enforced:
    • 3-20 characters
    • A-Z, a-z, 0-9, dash (-) are the ONLY acceptable characters
    • Space, underscore, punctuation, and special characters are NOT ALLOWED
    • No replication

Sample Labeling Conventions- General Guidelines

  • All tubes, plates, and boxes must include the following information.
    • PI name
    • Date
    • Sample name (tubes only)

Shipping or Delivering- General Guidelines

  • A manifest/service request must accompany all shipment or deliveries.
    • Schedule a time for drop off with a staff member at nextgen-seq@biotech.wisc.edu
    • Ship on dry or wet ice to:
      • DNA Sequencing Facility, 425 Henry Mall, 1250 Biotechnology, Madison WI 53706
  • For transport of biological samples on campus, please consult Biological Materials Transport on UW Campus Guidance. For shipment of biological materials, consult local and federal guidelines for safe packaging.
  • Samples delivered to us in an unsafe manner will either be refused or disposed immediately.

Whole Blood High Molecular Weight DNA Extraction - Specific Guidelines

  1. Follow ALL General Guidelines above.
  2. 1-5 ml anti-coagulated (EDTA – lavender top) whole blood. Please consult on smaller volumes.
    • ≥3 ml for ultra-long DNA extraction (only applicable to Oxford Nanopore ultra-long sequencing application).
  3. Submitted frozen in shatter-proof collection vessel (BD Vacutainer), 1.5-2 ml cryovial with external threads, or 1.5-2 ml snap-cap tube with tight-fitting lid.

Animal Tissues High Molecular Weight DNA Extraction - Specific Guidelines

  1. Follow ALL General Guidelines above.
  2. 25 mg total tissue weight.
  3. Submitted frozen in 1.5-2 ml cryovial with external threads or snap-cap tube with tight-fitting lid.

Plant Tissues High Molecular Weight DNA Extraction - Specific Guidelines

  1. Follow ALL General Guidelines above.
  2. 1-5 g of tissue. Fresh, newly expanded, etiolated leaves are highly preferred.
  3. Submitted frozen in 1.5 ml tube to 50 ml conical, depending on leaf size and shape.

Animal Cells High Molecular Weight DNA Extraction - Specific Guidelines

  1. Follow ALL General Guidelines above.
  2. 1E6 -5E6 cultured cells exclusive of medium, buffer, and cryoprotectant.
    • ≥6E6 cultured cells for ultra-long DNA extraction (only applicable to Oxford Nanopore ultra-long sequencing application).
  3. Submitted frozen in 1.5-2 ml cryovial with external threads or snap-cap tube with tight-fitting lid.

Bacterial Cells High Molecular Weight DNA Extraction - Specific Guidelines

  1. Follow ALL General Guidelines above.
  2. 1E9 – 5E9 cultured cells exclusive of medium, buffer, and cryoprotectant. Growing to mid-log phase (e.g. OD600 of 0.5-0.6 for E. coli) is highly preferred.
  3. Submitted frozen in 1.5-2 ml cryovial with external threads or 1.5-2 ml snap-cap tube with tight-fitting lid.