Method to amplify nucleic acids to generate fluorescence labeled fragments of conserved and arbitrary products

摘要

Disclosed herein are methods for the identification of the species, serotype, and strain of a microorganism. Also disclosed are primers for use in detecting such microorganisms and kits comprising such primers.

主权项

1. A method for the identification of the species, serotype, and strain of a microorganism comprising:
(a) amplifying DNA comprising variable sequences interspersed between highly conserved rDNA sequences by PCR and amplifying additional genomic sequences by random amplified polymorphic DNA (RAPD) PCR using a first primer of 13-14 bases in length and a second primer of 11-13 bases in length, said first primer comprising:
(i) 11-12 contiguous bases from a highly conserved 16S rDNA region;(ii) 2 bases at the 5′ end of said first primer that are not complementary to said highly conserved 16S rDNA region; and(iii) a fluorescent label; and (b) separating the amplified DNA produced in step (a).