The Flp double cross system a simple efficient procedure for cloning DNA fragments.

ABSTRACT:

Background

While conventional cloning methods using restriction enzymes and polynucleotide ligase are adequate for most DNAs, fragments made by the polymerase chain reaction are difficult to clone because the amplifying DNA polymerase tends to add untemplated nucleotides to the 3'-termini of the amplified strands. Conservative site-specific recombinases offer an efficient alternative to conventional cloning methods.

Results

In this paper I describe the use of the Flp site-specific recombinase for cloning PCR-amplified fragments. A DNA fragment is amplified with primers that contain at their ends inverted target sequences for Flp. Flp readily recombines these fragments in vitro into a vector that also contains two inverted Flp target sequences surrounding the alpha-complement

SUBMITTER: Sadowski PD 

PROVIDER: S-EPMC169169 | biostudies-literature | 2003 Jul

REPOSITORIES: biostudies-literature