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

{
  "publication_number": "US2013029341A1",
  "country": "US",
  "kind": "A1",
  "title": "Method to amplify nucleic acids to generate fluorescence labeled fragments of conserved and arbitrary products",
  "abstract": "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.",
  "claims": [
    "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 13-15 bases in length and a second primer of 11-13 bases in length, said first primer comprising: (i) at least 11 contiguous bases from a highly conserved 16S rDNA region; and (ii) a fluorescent label; and (b) separating the amplified DNA produced in step (a). 2. The method of claim 1, wherein said first primer is a forward primer. 3. The method of claim 2, wherein said first primer comprises SEQ ID NO:1. 4. The method of claim 3, wherein said first primer is SEQ ID NO:2. 5. The method of claim 1, wherein said second primer comprises 11-13 contiguous bases from 23S rDNA. 6. The method of claim 5, wherein said second primer is SEQ ID NO:3 or SEQ ID NO:4. 7. The method of claim 1, wherein said amplifying step utilizes a third primer of 11-13 bases in length, said third primer comprising 11-13 contiguous bases from 23S rDNA. 8. The method of claim 1, wherein step (b) is accomplished by capillary electrophoresis. 9. An isolated polynucleotide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and the full-length complements thereof. 10. A kit comprising a set of primers comprising PCR primers SEQ ID NO:2 labeled with a fluorophore and at least one of SEQ ID NO:3 and SEQ ID NO:4."
  ],
  "cpc": [
    "C12Q 1/689",
    "C12Q 1/6846",
    "C12Q 1/686"
  ],
  "assignees": [
    "AGARWAL ANJANA",
    "JENSEN MARK A"
  ],
  "filing_date": "2012-07-19",
  "publication_date": "2013-01-31",
  "priority_date": "2011-07-25",
  "application_number": "US-201213553256-A",
  "family_id": "46584402"
}