Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis

Abstract Cloning vectors derived from filamentous phage are extremely useful because they allow cloned DNA to be isolated as either single‐ or double‐stranded DNA. This unit contains protocols for preparing both forms of DNA and for characterizing inserts in M13‐derived vectors. A protocol is also presented for preparing single‐stranded DNA from plasmids using superinfection with helper phage. This method is advantageous because it allows cloned DNA to be maintained in the form of a plasmid while permitting single‐stranded DNA to be isolated for DNA sequencing.