The Research for the Extraction of Yeast'S Nucleic Acid with BMIM BF_4-H_2O-KH_2PO_4 Ionic Liquid Aqueous Two-Phase System

摘要

Construct ionic liquid - salt aqueous two-phase system for extracting yeast DNA. By using NaOH solution to process yeast, leaching the nucleic acid by ultrasonic assisting, variety of salts' aqueous two-phase influence on the ionic liquid 1-butyl-3-methylimidazolium fluoroborate are discussed, and the influence of additive quantity of KH_2PO_4 and PH on the ionic liquid aqueous two-phase system for extraction of nucleic acid are studied. The result shows that process the yeast with 0.3% NaOH solution, assisted by ultrasound with power of 80W, temperature 60°C, 3 hours' extraction. KH_2PO_4 take obvious advantage in the formation of ionic liquid aqueous two-phase system. In the process of extraction nucleic acid from [BMIM] BF_4-H_2O-KH_2PO_4 aqueous twophase process of nucleic acid, the best B-R buffer PH is 8.95, when the additive dosage of KH_2PO_4 is 2.000g, A260/A280 in the supernatant is smaller than control group, the substratum which rich in ionic liquid contains more nucleic acid than the supernatant, and A260 in the supernatant is more than that in the control group. Therefore, we conclude that the ionic liquid aqueous two-phase has a relatively good effect in extraction of nucleic.