In vitro transposition of artificial transposons for DNA sequenc

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435 6, 4351723, 435183, 4353201, 536 253, C12N 900, C12N 1566

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057285519

ABSTRACT:
We have developed efficient methods of creating artificial transposons and inserting these transposons into DNA targets in vitro, primarily for the purpose of mapping and sequencing DNA. A target DNA has been engineered to convert virtually any DNA sequence, or combination of sequences, into an artificial transposon; hence, custom transposons containing any desired feature can be easily designed and constructed. Such transposons are then efficiently inserted into DNA targets, in vitro, using the integrase activity present in yeast Ty1 virus-like particles. Primers complementary to the transposon termini can be used to sequence DNA flanking any transposon insertion.

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