Methods for the multiplexed isothermal amplification of nucleic acid sequences

A method for the surface bound clonal concatameric amplification of a plurality of nucleic acid sequences, comprising:

• (a) taking a population of different single-stranded nucleic acid molecules (ss1) of at least 50 nucleotides in length wherein the single strands have a different sequence and each of the single strands has a common hairpin region at the 3′ end (LP1) and a common hairpin region at the 5′ end (LP2), wherein each hairpin has 5′ and 3′ hybridising stem arms of between 15-30 base pairs in length, the stem arms having a Tm in the range of 45-55° C.;
• and
• (b) clonally amplifying the population of different single-stranded nucleic acid molecules (ss1) using a reaction mixture containing a strand displacing polymerase, nucleotide monomers and amplification primers wherein at least one of the amplification primers (P1) is complementary to at least a portion of the loop of LP1 and to at least a portion of the 5′ stem arm of LP1 and one of the amplification primers (P2) hybridizes to at least a portion of a copy of the loop of LP2 (LP2′),
• wherein in step (b) the amplification primer (P1) is attached to a solid support;
• and the population of single-stranded nucleic acid molecules are hybridized to the attached amplification primers (P1), such that single strands of different sequences are amplified in physical isolation;
• and the 3′ end of the stem arm of LP1 is extended to make a copy of the single-stranded nucleic acid molecules.