Promoter miR172c and application thereof
A technology in fragments and sequence lists, applied in applications, angiosperms/flowering plants, DNA/RNA fragments, etc., can solve the problems of gene expression accumulation at the outer edge and affect plant growth and development, and achieve the effect of strong specific expression activity
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[0024] Example 1. Preparation of promoter and verification of specific expression
[0025] 1. Preparation and confirmation of the promoter
[0026] 1. Extraction of rice genomic DNA
[0027] (1) Take the leaves of Nipponbare rice, add liquid nitrogen, after fully grinding, pour the powder into a 1.5ml centrifuge tube;
[0028] (2) After the liquid nitrogen in the centrifuge tube is completely volatilized, immediately add 500ul DNA extraction buffer (0.1mol / LTris HCl (pH 8.0), 0.5mol / L NaCl, 0.05mol / L EDTA, 0.5% SDS), fully After mixing, incubate at 65°C for 30 minutes, during which the centrifuge tube is gently inverted up and down to fully mix the sample with the buffer;
[0029] (3) Centrifuge at room temperature for 15 minutes at 12,000 rpm;
[0030] (4) Transfer the supernatant to another centrifuge tube;
[0031] (5) Add an equal volume of tris-phenol and extract once;
[0032] (6) Extract again with equal volume of chloroform;
[0033] (7) Take the supernatant, add...
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