Compatibility of DNA and RNA Extraction Methods for Challenging Plant Species

By: Eric Sonke3, Nezar Rghei1, Yousef Haj-Ahmad1,2 and Won-Sik Kim1, (1) Norgen Biotek Corp. 3430 Schmon Parkway, Thorold, Ontario, Canada, L2V 4Y6, (2) Brock University, 500 Glenridge Avenue, St. Catharines, ON, Canada, L2S 3A1, (3) Department of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, Western University London, ON, Canada.

Product No. E4788, E4913

Abstract

Frequent failure of downstream applications with DNA and RNA is often related to poor sample preparation. In particular, it is often difficult to extract DNA and RNA from samples that contain high levels of phenolic compounds, polysaccharides, volatile compounds and starch in an acceptable quality to be used in sensitive downstream applications such as PCR, RFLP and sequencing. Three different plant DNA and RNA isolation methods were validated to isolate genomic DNA and total RNA from challenging plant species, including raspberries, grapes, pears, pine needles and strawberry leaves. In the results, the relative yield of DNA and RNA, handling time and quality are systemically compared.

   

DNA Isolation



Proven DNA quality and quantity

Application: Random Amplification of Polymorphic DNA(RAPD) analysis

  • DNA was isolated from Prosopis cineraria using Norgen’s Plant/Fungi DNA isolation kit.
  • RAPD requires a high DNA quality to generate the band profile that discriminates amplification pattern for the species or sub-specie grouping.
  • 12 RAPD primers were successfully amplified the DNA fragments.

 

 

 

 


RNA Isolation



True compatibility and total RNA profile from challenging plant species



RNA Quality analysis using Bioanalyzer and spectrophotometer