Guidelines for collecting starting materials for Ultrapure and HMW DNA Extractions

All materials, regardless of organism, should be freshly-collected, immediately snap frozen in liquid nitrogen (or a dry ice ethanol bath), and stored at -80C until shipped on dry ice. For specific organisms, see details below.

Plants:

We have the best results starting with very young leaf/seedling tissue that has been grown in the dark for ~48 hours if possible, then snap frozen in liquid nitrogen at the time of collection. Using young tissue is important because it minimizes secondary compounds, minimizes waxy leaf coatings, minimizes presence of fungi or other growths on leaf surfaces, and offers the optimal ratio of cell weight to DNA content (i.e. more nuclei per cell weight).

The amount of plant material needed varies based on the type of plant as follows:

  • Dicot and Eudicot: ~50 grams
  • Monocot: ~60-90 grams

Animal tissue/cell lines:

Use ~2-3 grams of cells (or approximately 5x10e8 cells to 5x10e9 cells). Cells should be freshly collected then spun down (dump supernatant if cultured) and immediately snap frozen in liquid nitrogen. Frozen pellets can be stored at -80C until shipped on dry ice.

Bacterial or algae samples:

Use ~2-3 grams of cells (or approximately 5x10e8 cells to 5x10e9 cells). Cells should be freshly collected then spun down (dump supernatant if cultured) and immediately snap frozen in liquid nitrogen. Frozen pellets can be stored at -80C until shipped on dry ice.

Fungi:

Please provide ~5x10e8 protoplast cells or ~5-7 grams if counting is not possible. Important: Please do not freeze plugs or protoplasts.

Insects: 

We have the best results starting with ~3-5 grams of larvae (starved for 24-36 hours to minimize food in the gut), snap frozen in liquid nitrogen and stored at -80C until shipped on dry ice.

Deep sea organisms, soil samples, metagenomic samples:

Contact your sales representative or project coordinator at Amplicon Express for strategic details on collection. These organisms can be challenging to obtain high quality high molecular weight (HMW) DNA. Amplicon Express has optimized collection strategies and protocols for this purpose.

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