Internal reference genes suitable for researching flooding stress gene expression of lotus, and application thereof
An internal reference gene and gene expression technology, applied in the field of agricultural biology, can solve the problem of no internal reference gene, etc., and achieve the effect of improving stability
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[0028] 1. Material treatment: take lotus seeds as the material, soak the seeds under the conditions of 25°C and 16h / day light after the tail is broken, and select the seeds with the same bud length for the flooding stress test after 1-2 weeks after the emergence of hypocotyls. The flooding stress time was 1h, 2h, 4h, 12h.
[0029] 2. Fluorescence quantitative test: RNA extraction uses leaf bud material, the extraction method refers to the Shanghai Sangon column type plant total RNA extraction and purification kit manual (product number: B518661-0100), cDNA first strand synthesis refers to Shanghai Sangon M-MuLV First-Strand cDNA Synthesis Kit Instructions (Product No.: B532435-0020).
[0030] According to the five internal reference candidate sequences, primers were designed using primer premier5.0 software, and the primer sequences and estimated lengths of amplified fragments are shown in Table 1.
[0031] For the qPCR method, refer to the manual of Shanghai Sangon 2xSG Fast...
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