Specific molecular marker primer for genders of trachinotus ovatus, kit and application of kit
A technology of oval pomfret and molecular markers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problem of inability to identify the sex of oval pomfret live bodies, the inability to realize large-scale identification, Anatomy of individual death and other issues to achieve the effect of avoiding inbreeding depression, breaking through technical bottlenecks, and high accuracy
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Embodiment 1
[0030] The sex-specific molecular marker primers for ovoid pomfret provided in this example are primers P1, and the primers P1 include forward primers P1-F and reverse primers P1-R, and the forward primers P1-F The base sequence of the reverse primer P1-R is shown in SEQ ID NO:1, and the base sequence of the reverse primer P1-R is shown in SEQ ID NO:2.
[0031] Specifically, the base sequence of primer P1 is as follows:
[0032] Forward primer P1-F: 5'-AACTCTATGCACCGACCTGA-3';
[0033] Reverse primer P1-R: 5'-CATGAACAGACACGAGCACC-3'.
[0034] A full-sib family of Pompano ovata was collected, including 2 parents and 100 offspring individuals, and the physiological sex was identified after dissection. High-throughput sequencing was used to mark the parents and offspring with the genome sequence of the pomfret pomfret as a reference sequence, and linkage analysis was used to locate the sex quantitative trait loci and locate them in a single interval. Development of indels sign...
Embodiment 2
[0040] The method for male and female identification of oval pomfret provided by this embodiment comprises the following steps: using PCR amplification to amplify specific fragments in the genomic DNA of male and female individuals, specifically including: designing the primers in Example 1, according to the embodiment The method in 1 extracts the genomic DNA of the ovoid pomfret to be tested, adopts the primers in Example 1, and uses the extracted genomic DNA as a template to carry out PCR amplification, and adopts gel electrophoresis (specifically using an automatic sequencer ABI 3730XL) to detect The amplified products were identified as male and female individuals according to electrophoresis results.
[0041] The object of implementation in this embodiment is pomfret ovata, and 47 females and 47 males were taken, and the sexes were identified through anatomy, and then the feasibility of the primer was verified.
[0042] (1) Extraction of DNA
[0043] Cut about 1 square c...
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