Oil palm (Elaeis guineensis Jacq.) systems biology offers a comprehensive view of the plant system byemploying a holistic multi-omics approach encompassing the molecular data at various hierarchical levels.Sample limitation and the importance of integrating all molecular data with minimal variation, led to thedevelopment of sequential extraction of biomolecules fractions from a single undivided biological sample.Oil palm root was subjected to solvent extraction with methanol-chloroform-water to recover metabolitesof diverse polarity. The resultant pellet was subjected to buffer and solvent partitioning to obtain RNA andproteins. RNA extracted from the oil palm root showed a recovery of 180.25 ng mg^–1, with an A260:A280ratio ranging between 1.9-2.0 and RNA integrity number (RIN) value of 6.7. Co-extracted proteins resultedin a recovery of 29.28 µg per 100 mg fresh weight (FW) tissue and revealed a total of 1852 identifiedproteins in the oil palm root. Polar metabolites revealed approximately 40 metabolite peaks, and non-polarmetabolites with two major fatty acid groups i.e. saturated and unsaturated fatty acids at 55.4% and38.6%, respectively. This protocol demonstrated an advancement of extraction protocols for oil palm rootbiomolecules, which will consecutively expedite the establishment of various multi-omics platforms.