Purification of Amplifiable Nucleic Acid from Oil Palm and Rice Leaves and Seeds Using a Small Benchtop Automated Instrument

Plant nucleic acid (NA) purification provides a variety of unique challenges due to the broad diversity of plants, their extracellular structures, and endogenous compounds (e.g. polysaccharides, phenolics) that can co-purify with NA and inhibit downstream enzymatic assays.  We evaluated a novel cellulose-based paramagnetic particle technology to purify NA from Elaeis guineensis (oil palm) and Oryza sativa (rice) plant leaf and seed lysates in an automated format using the Maxwell® 16 System.  Extracted NA were evaluated for yield and amplifiability in qPCR and RT-qPCR.  When compared to a competing method, the Maxwell® 16 System isolated >10x more DNA from a single oil palm leaf punch, and unlike the competing method, showed no inhibition of subsequent qPCR when 2-4 leaf punches were used for extraction.  For leaf RNA and seed DNA, up to 100mg ground tissue were tested.  We were able to extract amplifiable NA in both cases with no detectable co-purification of inhibitors.  DNA extracted from oil palm leaves and seed was also amplifiable in endpoint PCR using SHELL gene-specific primers.  DNA from a single rice kernel was sufficient for genotyping by multiplex PCR.  We also tested DNA and RNA extraction from a variety of fruit and vegetable leaves including Malus domestica (apple), Vitis vinifera (grape), Citrus lemon (lemon), Solenum tuberosum (potato), and Solanum lycopersicum (tomato), and found all to yield amplifiable NA.  These data support the broad utility of the novel cellulose-based technology and Maxwell® 16 System for automated plant NA purification.