Molecular Marker and Application of Rice Glue Consistency Control Gene sbe1
A technology of molecular marking and gel consistency, which is applied in the field of agricultural biotechnology engineering, can solve problems such as poor accuracy, and achieve the effect of improving gel consistency
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Embodiment 1
[0060] Example 1. Using the Indel marker SBE1-m to identify the polymorphisms of the indica rice Teqing with high gel consistency and the japonica rice Nipponbare with low gel consistency;
[0061] The specific method is as follows: select the rice materials Nipponbare and Teqing, cross the Nipponbare and Teqing to obtain its F1, and use the primer SBE1-m to identify its polymorphism ( figure 1 ).
[0062] 1. DNA extraction
[0063] 1) Prepare DNA extraction buffer:
[0064] Add 1 volume of DNA extraction solution (0.35M sorbitol; 0.1M Tris, pH8.2; 0.005MEDTA; the rest is water), 1 volume of nuclear lysis solution (0.2M Tris, pH7.5; 0.05M EDTA; 2M NaCl; 0.055MCTAB; the rest is water) and 0.4 volume of 5% (mass concentration) sarkosyl solution (i.e. the aqueous solution of sodium lauryl-N-methylglycinate); finally add sodium bisulfite to prepare DNA extraction buffer solution; the final concentration of sodium bisulfite in DNA extraction buffer was 0.02M.
[0065] The prepa...
Embodiment 2
[0084] Example 2. Using the Indel molecular marker SBE1-m to identify sequence differences between the indica rice Teqing with high gel consistency and the japonica rice Nipponbare with low gel consistency
[0085] The specific method is: use the Indel molecular marker SBE1-m to perform PCR amplification on the genomic DNA of Nipponbare and Teqing, entrust Shanghai Yingjun Biotechnology Co., Ltd. to sequence the amplified products, and compare the differences in their sequences ( figure 2 ).
[0086] 1. DNA extraction
[0087] 1) Prepare DNA extraction buffer:
[0088] With embodiment 1.
[0089] 2), the paddy rice blade of above-mentioned Nipponbare and special green is carried out as follows respectively:
[0090] With embodiment 1.
[0091] 3) PCR amplification
[0092] With embodiment 1.
[0093] 4) Recovery of PCR products
[0094] The recovery of PCR products was carried out by using the PCR product recovery kit (spin column type, catalog number: DP1403) develope...
Embodiment 3
[0096] Example 3. Using the Indel marker SBE1-m to carry out assisted selection breeding for high gel consistency
[0097] The specific method is: Teqing, the gene donor parent with high gel consistency, is sequentially crossed, backcrossed and selfed with the japonica rice variety Nipponbare with low gel consistency, and the resulting progeny combined with molecular marker SBE1-m is assisted selection, and the segregation population is selected A single plant with the same belt type as the special green belt type is used for breeding improvement.
[0098] 1. DNA extraction
[0099] 1) Prepare DNA extraction buffer:
[0100] With embodiment 1.
[0101] 2), above-mentioned Nipponbare, Teqing, and the rice blades of the gained offspring are respectively processed as follows:
[0102] With embodiment 1.
[0103] 2. Indel marker detection
[0104] 1), PCR amplification
[0105] With embodiment 1.
[0106] 2), electrophoresis detection
[0107] With embodiment 1.
[0108] ...
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