Cold-tolerance gene qCT-3-2HHZ for rice booting stage stable expression and molecular marking method thereof
A stable expression, booting stage technology, applied in genetic engineering, biochemical equipment and methods, microbial assay/inspection, etc., to speed up the breeding process and facilitate hybridization.
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Embodiment 1
[0021] (1) Mapping of cold tolerance genes
[0022] 1. Test materials
[0023] Huang Huazhan, IR64PSBRC28, PSBRC66, IR50, Phalguna, OM1723, CDR2, Teqing, Huang Huazhan and the latter 8 parents (donor parents) are interrelated breeding populations. The population consists of 497 lines composed of 8 families .
[0024] 2. Sequencing and SNP screening
[0025] DNA extraction of 9 parents and populations was performed using DNeasyPlantMiniKit (Qiagen) kit. Resequencing and sequence alignment were performed by Huada Gene Technology Co., Ltd. With minimum allele frequency (MIF) > 0.05 and missing data < 10% as screening criteria, our research group used SAMtools to complete high-quality and high-density SNP screening and implement genotype analysis of test materials.
[0026] 3. Genome-wide association analysis
[0027] First input the SNP genotype data of the population into the mapping software (TASSEL5.2), evaluate the linkage disequilibrium value (LD value) and use the Struct...
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