Transposon nucleic acids comprising a calibration sequence for DNA sequencing

摘要

Transposon nucleic acids comprising a transposon end sequence and a calibration sequence for DNA sequencing in the transposon end sequence. In one embodiment, the transposon end sequence is a Mu transposon end. A method for the generation of DNA fragmentation library based on a transposition reaction in the presence of a transposon end with the calibration sequence providing facilitated downstream handling of the produced DNA fragments, e.g., in the generation of sequencing templates.

主权项

1. An in vitro method for fragmenting target nucleic acids comprising contacting the target nucleic acids with a plurality of MuA transposon complexes, under a condition for transposon-mediated nucleic acid fragmentation, to generate a plurality of nucleic acid fragment products having a calibration sequence added to one end, wherein the MuA transposon complexes contain a first and a second nucleic acid duplex and a plurality of MuA transposon proteins, wherein the first and the second nucleic acid duplexes contain (i) a first strand having the nucleotide sequence of SEQ ID NO:1 that is modified to include a calibration sequence near the 3′ end, and (ii) a second strand that is fully complementary to the first strand, wherein the calibration sequence is four nucleotides in length and contains any nucleotide bases A, T, C, and/or G in any order and the calibration sequence replaces any four nucleotides in the first strand near the 3′ end in a region located at position 39-50 of SEQ ID NO:1.