Nitrate reductase gene BcNR of Chinese cabbage
A technology of head cabbage and reductase, which is applied in the field of molecular biology and biology, and can solve problems such as difficult to obtain satisfactory results
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[0027] (1) Cloning of BcNR gene cDNA
[0028] The non-heading Chinese cabbage variety 'Suzhouqing' (a local variety in Jiangsu, purchased from the market) was selected as the material. After the cotyledons were flattened, total RNA was extracted from the leaves according to the instructions of the Simply P Total RNA extraction kit from Bioflux Company, and 2 μg of total RNA was taken to synthesize cDNA. The cDNA product was digested with RNase.
[0029] According to the conserved nucleotide sequences in Arabidopsis NIAI and NR Brassica napus, design three pairs of specific primers NRF1 / NRR1: SEQ ID NO.3 / SEQ ID NO.4, NRF2 / NRR2: SEQ ID NO.5 / SEQ ID NO.6, NRF3 / NRR3: SEQ ID NO.7 / SEQ ID NO.8, use LA Taq enzyme to carry out conventional polymerase chain reaction (PCR), amplify the BcNR gene segmentally, the amplification program is: 94°C Pre-denaturation for 5 minutes, denaturation at 94°C for 30 sec, renaturation at 55°C for 30 sec, extension at 72°C for 1 min and 30 sec, after 35 ...
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