Method for screening medium amylase content rice and primer special for method
An amylose content, rice technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of inability to separate medium amylose and high amylose genotypes, etc. Breeding cycle, short cycle, cost reduction effect
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Embodiment 1
[0018] Using the special primer Wx-Ex6 in the present invention, molecular detection was carried out on 200 micro-core collections of cultivated rice in China and 95 core collections of foreign introduced species, including 111 indica varieties and 89 japonica varieties.
[0019] DNA was extracted using conventional CTAB extraction method.
[0020] The PCR amplification system is: rice genomic DNA template 50ng, Taq DNA polymerase 1 U, 1.5μl 10×PCR buffer (100Mm Tris-HCl (pH8.3), 500Mm KCl), 25mM MgCl 2 0.9ul, 2.5 mmol each dNTPs 0.6ul, 0.2μM for each of the four primers, and finally use ddH 2 O Supplement the reaction system to 15 μl.
[0021] The PCR reaction conditions can be: first 95°C for 5min; then 95°C for 30sec, 55°C for 30sec, 72°C for 40sec, a total of 35 cycles; then 72°C for 5min.
[0022] Detection of PCR products: Load the PCR products on a 1.5% agarose gel to detect the band pattern of the PCR products.
[0023] Molecular results showed that there were 766b...
Embodiment 2
[0026] First plant the above-mentioned Jufeng A / / Tianfeng A / J521 F in the field 6There are 114 lines in total in the generation breeding group; J521 contains the pedigree of the medium-straight variety Lemont, while Jufeng and Tianfeng are both tall-straight varieties. The method of the invention and the provided molecular marker Wx-Ex6 are used for detection at the seedling stage. DNA extraction, PCR amplification system and PCR product detection PCR product detection methods are the same as in Example 1.
[0027] DNA was extracted from these 114 strains, and the above-mentioned detection process was carried out, such as figure 1 As shown, the leftmost band is the molecular weight control Marker DL2000, P1 is the band type of the positive parent Lemont, the band size is 1025 and 766bp; P2 is the band type of the negative parent Jufeng A, the band size is 1025 and 310bp; 1-20 are the 20 rice lines to be detected, and the ones showing the same band size as P1 are wxya in P...
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