A genetic molecular marker and application for fast-growing sea cucumber breeding
A molecular marker, sea cucumber technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Efficient, targeted results
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Embodiment 1
[0022] Example 1: Acquisition of genetic molecular markers for breeding of sea cucumber growth traits and their verification in the offspring of the family
[0023] A sea cucumber family was constructed, and the 2 parents and 142 offspring of the family were used as the mapping population. With the help of a high-throughput sequencing platform, SLAF markers were developed, and a high-density genetic linkage map of sea cucumber was constructed using HighMap software. After high-throughput sequencing, a total of 264,810 SLAF tags were developed, including 112,322 polymorphic SLAF tags. The average sequencing depth of parents with SLAF tags was 23.67×, and the average sequencing depth of offspring was 5.44×. Through bioinformatics analysis, there are 50,905 tags that can be used for genetic map construction. After quality filtering, 4629 polymorphic tags with high sequencing depth, high integrity and no partial separation were obtained. The screened 4629 SLAF tags were used for ...
Embodiment 2
[0027] Example 2: Verification of Apostichopus japonicus growth-related SNP molecular markers in an expanded population
[0028] According to the flanking sequences on both sides of the SNP160 site obtained by high-throughput sequencing, primers for SNP site amplification detection were designed, and the corresponding primer sequences were SNP160F:5'-TCGCAGGAAACCAACCTTCA-3' and SNP160R:5'-AAGGGAGGGTCAAGGAGGAT-3' . The corresponding sequence was used to type the SNP site in the expanded population by using the HRM small fragment method, and the QTL site verification was carried out in combination with the related trait data of the expanded population. The expanded population used was 10-month-old seedlings from the same batch of large-scale breeding and cultured in the same breeding environment. 106 individuals were randomly selected, and the body weight of the individuals was measured. The genotype detection of individuals is carried out by the HRM method. The amplification d...
Embodiment 3
[0032] Example 3: Application of molecular markers for growth traits-assisted selection breeding in Apostichopus japonicus breeding and production
[0033] During the spring seedling period of A. japonicus, 100 parent references were randomly selected from the cultivated parent references, and the genotypes of 100 reference references at the SNP160 locus were genotyped using HRM technology. The primer sequences used were SNP160F: 5'-TCGCAGGAAACCAACCTTCA-3' and SNP160R: 5'-AAGGGAGGGTCAAGGAGGAT-3', and the amplification detection system was: DNA template 1μL (50ng / μL), 4.5μL 2×ES Taq Master Mix, 0.5 μL (10 μmol / L) each of upstream and downstream primers, 3.5 μL ddH 2 O; PCR reaction program: pre-denaturation at 95°C for 5min; denaturation at 94°C for 30s, annealing at 59.5°C for 30s, extension at 72°C for 30s, a total of 35 cycles; final extension at 72°C for 7min, and storage at 4°C; Melting curve analysis technique (HRM) method was carried out. According to the typing results,...
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