Method for detecting three-line hybrid rice honglian-type new restoring gene by using molecule mark sensing
A technology for three-line hybrid rice and gene restoration, which is applied in the field of molecular marker detection, can solve problems such as differences in the relationship between restoration and conservation, and the inability to restore CMS-D2-2, etc., to achieve the advantages of convenient operation, increased genetic diversity, and broadened the range of selection Effect
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[0060] Using PCR amplification to quickly detect the molecular markers of the rice red lotus type new restorer gene, combined with genetic analysis to screen the red lotus red lotus type new restorer gene as an example, the specific steps are as follows:
[0061] 1. PCR primer design
[0062] The primers were designed based on the co-segregated molecular markers M1 and M2 of the rice red lotus-type cytoplasmic restorer genes Rf5 and Rf6. The sequences are as follows:
[0063] M1 primer:
[0064] Forward primer F1: 5'-ATGACAAATCTGCTCCGATG-3'
[0065] Reverse primer R1: 5'-CTTACTTAGGAAAGACTAC-3'
[0066] M2 primers:
[0067] Forward primer F2: 5'-GGAGATGCTATAGCAGCAGTG-3'
[0068] Reverse primer R2: 5'-ATTGCTCCTTACCACCTTGC-3'
[0069] 2. Extraction of DNA:
[0070] For each rice variety, take a section of rice tender leaves with a length of 3-5cm, a width of 0.5-1cm, and a weight of about 0.1g and 0.2g of powdered quartz sand, put them in a 1.5ml centrifuge tube in turn, gr...
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