Sample Preservation Protocol RNA/DNA Shield

Protocol for the preservation of animal tissues for RNA and DNA extraction in the absence of liquid nitrogen and -80 freezers (i.e., remote field collection protocol)

Necessities

Reagents

Equipment

RNALater Guidelines

RNA/DNA Shield

  • Use RNA/DNA Sheild Solution with fresh tissue only; do not freeze tissues before immersion in RNAlater Solution.
  • Before immersion in Solution, cut large tissue samples to ≤0.5 cm in any single dimension.
  • Place the fresh tissue in shield following manufacturer’s instructions.

Sample Collection

Tool Sterilzation

  • Make 10% bleach solution in DI water in plastic beaker labeled #1 10% Bleach
  • Pour 100% DI water in plastic beaker labeled #2 DI Water
  • Pour 100% Ethanol soluion in plastic beaker labeled #3 100% Ethanol

  • Line up Beakers 1, 2, and 3 next to the ethanol burner filled with 100% ethanol

  • First, dip the clippers and forceps in #1 10% Bleach
  • Second, rinse clippers and forceps in #2 DI Water
  • Third, rinse the clippers and forceps in #3 100% Ethanol
  • Fourth, sterilze tools by burning off the ethanol
  • Fifth, place tools upside down in tube rack with sterile end up

Glove Cleaning

  • Make a 5 gallon bucket with 10% bleach
  • Make a 5 gallon bucket with fresh water
  • Dunk gloved hands in 10% bleach and then fresh water bucket between each sample

Sample Collection

  • Fill each tube with desired volume of RNA DNA Shielf solution
  • Pick up sample with clean glove
  • Cut tissue section ≤0.5 cm in any single dimension
  • Place tissue section in prelabeled 2ml tube with Shield solution
  • Repeat sample collection process for a duplicate tube for every sample as possible

Sample Preservation

  • Store and/or transport samples as follows in DNA/RNA Shield for later purification of high-quality DNA and/or RNA
Temperature Time
4°C - 25°C (ambient) Up to 30 days
37°C Up to 3 days
-20°C and below >1 year
Written on June 1, 2016