Screening method and application of SSR loci for paternity testing
A screening method and paternity testing technology, applied in the field of biology, can solve problems such as breeding deviation, economic loss, and unknown pedigree records, and achieve the effects of high accuracy, high screening efficiency, and a wide range of applications
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Embodiment 1
[0084] The present invention will be further explained below by taking the screening of SSR molecular markers for yak paternity identification as an example.
[0085] (1) Public data selection
[0086] Public data selection: From the NCBI SRA database, screen the whole genome sequencing data using yak genomic DNA sequencing as the data source, including whole genome sequencing projects and resequencing projects, a total of 114 samples, 126 Runs (one in the sequencing process) . In order to avoid the specificity of short sequence fragment recognition between different platforms, a total of 75 sequencing samples with the same platform-Hiseq2000 sequencing platform and uniform sequencing data quality were randomly selected.
[0087] (2) Data calibration and data collation
[0088] Data calibration and collation: the collected SRA data is first converted to FASTQ data format with the "fastq-dump-split-3" command of sra-tool kit (v2.5.2), and cutadapt (v.1.8.1) program (Default ...
Embodiment 2
[0177] Sample collection: Blood samples from live yaks were collected from the jugular vein, added with blood anticoagulant EDTA, and stored in a -80°C ultra-low temperature refrigerator. The samples are positive samples with clear records. There are 6 families with a total of 18 heads (samples include father, mother, and offspring) from the Longri species storage farm in Sichuan Province. These samples are used for parental analysis and clinical testing.
[0178]从筛选出的50个SSR位点中选择如下20个SSR位点,SSR geneID为scaffold2072_1、scaffold341_1、scaffold1139_1、scaffold879_1、 scaffold2036_1、scaffold2058_1、scaffold1000_1、scaffold1649_1、 scaffold4112_1、scaffold506_1、scaffold1645_1、scaffold2687_1、 scaffold1210_1、scaffold547_1、scaffold1214_1 , scaffold1343_2, scaffold1141_1, scaffold94_1, scaffold738_1, and scaffold629_1.
[0179] Design of microsatellite primers: Design PCR amplification primers for 20 yak paternity identification microsatellite loci, and modify the 5' end of each forward primer w...
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