A Modified Method of Total RNA Isolation for Mango Leaf Tissues

Abstract

Complex metabolic components in mango leaves lead to high difficulty in RNA extraction. In order to improve the quality of total RNA extraction, on the basis of the existing methods reported in the literature, an RNA extraction method combining acetone washing liquid nitrogen abrasive material with 0.3 mol/L lithium chloride (LiCl) solution and 2 times the volume of anhydrous ethanol as the second precipitator was developed. The experimental results showed that the improved method significantly reduced the amount of impurity precipitation in the RNA extraction process. The electrophoretic bands of total RNA extracted were complete. The absorbance ratio of OD 260/280  was about 1.9, and the average was 329.8 g/g FW±11.2 g/g FW. Further validation experiments of RNA reverse transcription and the polymorphic amplification of double-stranded cDNA related sequences (RAP-PCR) obtained clear polymorphic bands, showing that the total RNA from mango leaves had high quality and was suitable for molecular biology experiments.