FastPrep® and associated matrices have demonstrated successful lysis and RNA extraction from cassava roots in only 60 seconds..

Introduction

Because plant samples can be very fibrous and contain high levels of polyphenolic compounds, polysaccharides, and RNases, it can be extremely difficult to extract enough usable DNA or RNA for PCR analysis and other downstream applications. The FastDNA™ Kit, FastDNA™ Spin Kit, and FastRNA™ Pro Green Kit make it fast and simple! Effective, efficient sample preparation is critical to successful downstream results.

Overview

  • Keywords: RNA extraction, roots, fibrous plants
  • Aim of the study: Optimization of RNA extraction from fibrous roots
  • Application: RT-qPCR
  • Sample name: Cassava Root
  • Sample type: Root
  • Material: FastPrep-24™ instrument, Lysing Matrix A with an additional 1/4 –inch ceramic bead

Protocols & Parameters

  1. Add the Cassava root sample to a Lysing Matrix A tube containing an additional 1/4-inch ceramic bead.
  2. Add 1 ml of RNApro extraction buffer.
  3. Homogenize in the FastPrep-24™ instrument for 60 sec at a speed setting of 6.0.
  4. Centrifuge at 14,000 x g for 5–10 minutes to pellet debris.
  5. Collect the supernatant and proceed with the RNA extraction protocol.

Results


RNA extraction from Cassava storage roots; samples contain 0.32 ųg/ųl–110 ųg/ųl RNA. FastPrep settings: Speed 6.0 for 60 s; Lysing Matrix A with additional ceramic 1/4-inch bead.

Conclusion

  • RNA could successfully be extracted from cassava roots with the FastPrep® System. Total RNA yields achieved were up to 110 μg of RNA per μl.