US 6,593,113 B1 · In vitro method for providing templates for DNA sequencing

Abstract

The present invention relates to an in vitro method for providing unique templates for DNA sequencing and to a kit, which can be used, when employing the selection method of this invention. The examined DNA is subjected to a DNA transposition reaction and to an amplification reaction. The amplification reaction is carried out in the presence of a fixed primer and a selective primer having a complementary sequence to the end of a transposon DNA. A transposition reaction can be carried out also in the presence of a transposon or transposons, which have different ends enabling the design of selective primers for both ends. Each size of the amplification products represents one template for a sequencing reaction.

Claims14

Dependent claims

An in vitro method for providing templates for DNA sequencing, comprising the steps of

carrying out a transposition reaction in the presence of a target DNA and in the presence of a transposon, said transposition reaction resulting in at least one insertion into said target DNA;
performing an amplification reaction by using as primers a first, fixed primer hybridizing at a known location in said target DNA or, if the target DNA is part of a vector, adjacent to said target DNA, and a second, selective primer hybridizing at the transposon insertion site, said selective primer having a sequence complementary to the joining end of said transposon and having additional nucleotides of known identity at its 3′ end;
separating the amplification products on the basis of their size, and
recovering said amplification products suitable for use as templates in sequencing said target DNA.

1 more element

Dependencies are inferred from the claim text.

In vitro method for providing templates for DNA sequencing

Life of this patent

In vitro method for providing templates for DNA sequencing

Life of this patent

Abstract

Claims14