Deprotection of oligonucleotides and purification using denaturing PAGE.
The advantages of purification by denaturing polyacrylamide gel electrophoresis (PAGE) are speed, simplicity, and high resolution. Although yields tend to be low (<50% of applied sample), the amount of material recovered is usually far in excess of that required for most molecular biology applications (e.g., cloning and sequencing). Denaturing PAGE can resolve oligonucleotides from 2 to 300 bases, depending on the percentage of polyacrylamide used. The method presented here is thus useful not only for isolating chemically synthesized deoxyribonucleotides but also small RNAs or other single-stranded polynucleotides.
Ellington
Andrew
A
Indiana University, Bloomington, Indiana.
Chory
Joanne
J
The Salk Institute, La Jolla, California.
D003001:Cloning, Molecular / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA / Q000379:methods
D009841:Oligonucleotides