STR (short tandem repeat) fluorescent multiplex amplification kit for pigeon individual recognition

Abstract

The invention belongs to the technical field of gene detection, and particularly discloses an STR fluorescent multiplex amplification detection kit for pigeon individual recognition, a detection method and application of the STR fluorescent multiplex amplification detection kit. The kit comprises 19 STR (short tandem repeat) loci and one sex identification locus of the domestic pigeon, also comprises an allele lader and the like, can more accurately judge the typing of the loci, has high cumulative paternity exclusion rate, can be applied to individual identification and paternity relationship identification of the domestic pigeon, has high detection sensitivity, and can obtain an accurate typing result only by 0.5 ng of DNA (deoxyribonucleic acid).

Claims

1. A STR fluorescent composite amplification detection primer composition for individual identification of domestic pigeons, characterized in that: The primers are directed to the STR loci and sex identification loci of domestic pigeons.

2. The STR fluorescent composite amplification detection primer composition according to claim 1, characterized in that: The STR loci are: CliμD11, PIGN15, PG1, PIGN10, CliμT43, CliμD17, PIGN57, CliμT13, PIGN04, PIGN26, CliμD01, PG6, CliμD19, PIGN12, CliμT02, CliμD32, CliμD35, CliμT17 and CliμD16; the gender identification locus is CHD.

3. The STR fluorescent composite amplification detection primer composition according to any one of claims 1-2, characterized in that: The primer sequence is as shown in SEQ ID NO. 1-40; or has at least 98% homology to SEQ ID NO. 1-40.

4. The STR fluorescent composite amplification detection primer composition according to any one of claims 1 to 3, characterized in that: The primer compositions are divided into 4 groups: Group 1: CliμD11, PIGN15, PG1, PIGN10, CliuT43; Group II: CliμD17, PIGN57, CliμT13, PIGN04, PIGN26; Group 3: CliμD01, PG6, ClimD19, PIGN12; Group 4: CliμT02, CliμD32, CliμD3, CliμT17, CliμD16, CHD.

5. The STR fluorescent composite amplification detection primer composition according to claim 4, characterized in that: The forward primers in the primers are all fluorescently labeled; preferably, the fluorescent labels are located at the 5' ends of the primers, and each group is labeled with 4 different fluorescent markers; more preferably, the 4 different fluorescent markers are FAM, HEX, TAMRA and ROX, respectively.

6. A STR fluorescence composite amplification detection kit for individual identification of domestic pigeons, characterized in that: The invention comprises the STR fluorescent composite amplification detection primer composition described in any one of claims 1 to 5.

7. The STR fluorescence composite amplification detection kit according to claim 6, characterized in that: The kit further comprises PCR Master MIX, an allele typing standard and a molecular weight internal standard; preferably, the allele typing standard comprises a four-base repeat sequence and a two-base repeat sequence; and the molecular weight internal standard is labeled with a fifth fluorescent ORG.

8. Use of the STR fluorescent composite amplification detection primer composition according to any one of claims 1 to 5 in individual identification and / or parentage testing of domestic pigeons.

9. Use of the STR fluorescent composite amplification detection primer composition according to any one of claims 1 to 5 in preparing a kit for domestic pigeon individual identification and / or parentage testing.

10. A method for individual identification and / or parentage testing of domestic pigeons, characterized in that: The step of amplifying and detecting a pigeon DNA sample using the STR fluorescent composite amplification detection primer composition described in any one of claims 1-5 or the kit described in any one of claims 6-7.

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