Molecular marker for early identification of sex of A. arguta young seedlings and application of molecular marker
A technology of kiwifruit and molecular markers, which is applied in the field of early identification of molecular markers for the sex of kiwifruit jujube seedlings, can solve the problems of high labor and time cost, poor versatility, no gender specificity, etc., and achieves good application prospects and stability. Good, universal effect
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Embodiment 1
[0044] Example 1: Development of Actinidia jujube-specific sex marker primers
[0045] (1) Using 〔Ruby Star (Actinidia jujube variety)×11-17 (Male Jujube)] hybrid seed population with 15 male and female plants plus 2 male and female plant parents, extract DNA to establish a BSA mixed pool; apply Illumina HiSeq sequencing technology The platform performs sequencing and bioinformatics analysis, including genome assembly and reference genome comparison statistics, SNP detection and annotation, Small InDel detection and annotation, sex-determining gene association analysis and primer sequence development.
[0046] A total of 31Gbp Raw Data was obtained by sequencing, and the Clean Data obtained after filtering was 30Gbp, with Q30 reaching 92.67%, and the average sequencing depth of each individual was 15X. The average comparison rate between the sample and the reference genome was 87.42%, the average coverage depth was 15X, and the genome coverage was 74.81% (at least 1X coverage)...
Embodiment 2
[0064] Example 2: Early sex identification method of Actinidia jujube
[0065] (1) Extraction of Genomic DNA from Actinidia jujube leaves
[0066] (1) Sampling and storage: young leaves of Actinidia jujube were taken, frozen in liquid nitrogen, and stored in a -80°C refrigerator.
[0067] (2) Extraction and preservation of leaf genomic DNA: take 100-200 mg of frozen leaf tissue, put it into a 2ml EP tube with 1 steel ball, freeze it again in liquid nitrogen, and grind it into a fine powder with a mill. Using the Plant Genomic DNA Rapid Extraction Kit (full name: DH08CTAB Plant Genomic DNA Rapid Extraction Kit) produced by Beijing Pulehai Biotechnology Co., Ltd., the CTAB method was used to extract the genomic DNA of the sample, and the specific extraction steps were carried out according to the kit instructions. The template DNA concentration was diluted to 20ng / μL, and stored in a -20°C refrigerator for later use.
[0068] (3) Main instrument models:
[0069] The centrifug...
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