Sulfonation of polyvinylidene difluoride resin and its application in extraction of restriction enzymes from DNA digestion solutions.
Sulfonation of polyvinylidene difluoride (PVDF) resin was achieved by incubation of the resin with sulfuric acid at a moderately high temperature. The sulfonated PVDF (SPVDF) resin was studied for its ability to extract restriction enzymes from DNA digestion solutions. The SPVDF resin was effective in adsorbing restriction enzymes such as EcoRI and BamHI and the extraction procedure was easy and simple to perform. The adsorption depended upon the amount of the resin added. We found that 1 mg of the SPVDF resin could completely remove all restriction enzyme activity routinely used in DNA digestion within 2 min after its addition. Treatment of a digestion solution with the SPVDF resin did not change the reaction solution and the same digestion buffer could be used for another digestion of the same DNA with other enzymes. We also found that, in comparison with normal PVDF, the SPVDF resin adsorbed less DNA, resulting in less loss of DNA in the extraction step. The potential application of the SPVDF resin in other procedures of molecular cloning and enzyme purification is discussed.
Yang
Jing
J
College of Life Sciences, State Key Lab of Protein and Plant Genetic Engineering, Peking University, Beijing 100871, China.
Dong
Chunxia
C
Huang
Xu
X
Zhao
Jindong
J
D000042:Absorption; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry*; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D011145:Polyvinyls / Q000737:chemistry*; D013464:Sulfuric Acids / Q000737:chemistry; D005465:Fluorocarbon Polymers
D004247:DNA; D004262:DNA Restriction Enzymes; D011145:Polyvinyls; D013464:Sulfuric Acids; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C033158:sulfuric acid