DNAgard® Protocol

Product Nos. 63020-016, 62001-036, and 62001-046

Product Description

DNAgard Tissue is designed for room temperature storage and shipment of DNA in biological tissues, such as mammalian cells and organ tissues. DNA in complex samples is protected by the unique stabilization properties of DNAgard Tissue. Samples can be stored in liquid DNAgard Tissue for at least 6 months at room temperature. Samples stored in liquid DNAgard Tissue are ready for immediate processing for DNA recovery via column extraction (following manufacturer’s instructions) or using standard lab procedures involving digestion and organic extraction. Purified DNA can be used directly in downstream applications.

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

Storage/Stability

DNAgard Tissue must be stored at room temperature. Use within 6 months of purchase date for optimal product performance. DNAgard Tissue stabilizes genomic DNA in cultured cells and animal tissue samples for at least 6 months at room temperature in a liquid storage format.

Procedures

Sample Storage

Liquid Storage of Tissue Samples in DNAgard Tissue

  1. Prepare tissue samples by dissection. For optimal DNA protection, store tissue fragments less than 75 mg. Small tissue fragments, thinly sliced (at least one edge of the tissue fragment be 5mm or less in length), ensures that DNAgard Tissue permeates rapidly the entire tissue sample. To maintain the integrity of the DNA, tissue fragments should be kept cold during dissection and transferred to DNAgard Tissue as soon as possible.
  2. Submerge tissue fragment in 500 µl DNAgard Tissue solution for shipment or storage. At least 100 µl DNAgard Tissue is required per 10 mg tissue. If sample is to be shipped, it is important to select a tube size that ensures that the tissue remains submerged during handling. We recommend the use of screw-cap tubes to prevent sample leakage during transport. For general storage, use 2 ml screw-cap tubes.
  3. Store samples at room temperature and protected from light for up to 6 months.

 

Notes on Sample Recovery (from tissue samples stored as liquid in DNAgard Tissue)

Processing samples for DNA recovery should be done by commercially available column purification technologies or via standard laboratory procedures involving tissue lyses and organic extraction. For ease of use, we recommend removing the DNAgard Tissue solution from the tissue fragment prior to DNA isolation. However, if DNA yield is critical, optimal DNA recovery can be achieved by isolating genomic DNA from the entire DNAgard Tissue sample.  Note on tissue disruption: DNA isolation via commercially available column technologies can often be facilitated by disruption of the tissue sample, thereby reducing the time required to fully lyse the tissue and release genomic DNA. We recommend the use of a pestle. Consult the DNA isolation manufacturer’s instructions.

Maximal recovery of genomic DNA. For column purification protocols allowing DNA isolation from tissue samples re-suspended in buffer, no additional processing of DNAgard Tissue samples is needed. Follow manufacturer’s instructions for DNA isolation, adhering to buffer-to-sample ratio specifications.

Maximal recovery of genomic DNA (modified protocol for column purification kits that initiate from a tissue sample free of liquid). Simply add the kit’s initial lyses buffer in a 1:1 ratio with the DNAgard Tissue volume. Scale all other reagents as necessary based on this initial volume (proceed as if the resultant mixture was entirely kit lysis buffer) and process according to the kit specifications.

Maximal DNA isolation involving organic extraction. Tissues stored in DNAgard Tissue can be processed for DNA isolation using standard lab protocols involving tissue lyses and organic extraction. Simply pipette off the DNAgard Tissue solution, being careful to not remove the tissue sample. Add lyses buffers and enzymes according your established protocol.

 

Notes on Sample Recovery (from tissue culture cells stored as liquid in DNAgard Tissue)

Samples can be processed for DNA recovery directly via commercially available column purification technologies – no additional processing is needed. DNAgard Tissue solution should not be removed prior to sample processing; add lyses buffers and enzymes provided in your DNA isolation kit directly to the DNAgard Tissue sample. Follow manufacturer’s instructions for DNA isolation, adhering to buffer-to-sample ratio specifications. Do not use organic extraction methods for DNA isolation (i.e. phenolchloroform extraction).

Maximal recovery of genomic DNA (using a DNA isolation kit that specifies cells be re-suspended in buffer or media). DNAgard Tissue can be treated as if it were any re-suspension buffer or media. Follow manufacturer’s instructions for DNA isolation, adhering to reagent ratio specifications.

Maximal recovery of genomic DNA (using a DNA isolation kit that only provides protocol from cell pellet starting point). Do not pellet the DNAgard Tissue-cell suspension (genomic DNA from cells is released into the DNAgard Tissue solution during storage). Simply add the kit’s initial lysis buffer in a 1:1 ratio with the DNAgard Tissue volume. Scale all other reagents as necessary based on this initial volume (proceed as if the resultant mixture was entirely kit lyses buffer) and process according to the specifications described in the kit.

Materials

     
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