Deproteination of nucleic acids by filtration through a hydrophobic membrane.
To remove or inactivate an enzyme from DNA in multistep procedures in molecular biology, it is often necessary to phenol extract the solution, followed by chloroform extraction and ethanol precipitation. In addition to being time-consuming and hazardous, there can be significant loss of DNA with this procedure, especially when small volumes or amounts of DNA are being used. We have found that filtering analytical reaction mixtures through a hydrophobic membrane specifically to remove protein is a rapid alternative to phenol extraction. Within broad limits commonly encountered in molecular biology, filtration through a polyvinylidene difluoride membrane quantitatively removes a variety of enzymes without significant loss of double-stranded nucleic acid.
Harley
C B
CB
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
Vaziri
H
H
D000469:Alkaline Phosphatase; D001665:Binding Sites; D004247:DNA / Q000302:isolation & purification*; D004277:DNA, Single-Stranded; D004798:Enzymes / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods; D008567:Membranes, Artificial; D009841:Oligonucleotides / Q000032:analysis; D011145:Polyvinyls; D012313:RNA / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic; D005465:Fluorocarbon Polymers
D000469:Alkaline Phosphatase; D004247:DNA; D004277:DNA, Single-Stranded; D004798:Enzymes; D008567:Membranes, Artificial; D009841:Oligonucleotides; D011145:Polyvinyls; D012313:RNA; D012259:Ribonuclease, Pancreatic; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers