Cloning Method of Cytochrome Oxidase Gene of Peri. didentatus
A technology of Nereis didentate and cytochrome, which is applied in the field of genetic engineering, can solve the problems that affect the wide application of Nereis didentate and the inability to study the gene of cytochrome oxidase in Nereis didentate, and achieve a cost-effective Effect
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[0028] The cloning method of the cytochrome oxidase gene of the bidentate silkworm is as follows:
[0029] One. Extract and purify the total RNA of the bidentate silkworm, synthesize the first strand of cDNA;
[0030] 1. Extraction and purification of total RNA: Use RNAiso Pius from Bao Bioengineering (Dalian) Co., Ltd. The extracted and purified total RNA electropherogram is as follows figure 1 Shown. Among them, M is a molecular weight marker, 1, 2, and 3 are purified total RNA.
[0031] 2. Synthesis of the first strand of cDNA: Use the RNA PCR Kit (AMV) Ver.3.0 kit of Bao Biological Company to mix the following reagents:
[0032] Mgcl 2 2 μl
[0033] 10×Reverse transcription buffer 1 μl
[0034] Rnase Free dH 2 O 3.75 μl
[0035] dNTP 1 μl
[0036] RNase inhibitor 0.25 μl
[0037] AMV reverse transcriptase 0.5 μl
[0038] Adapter primer Oligo dT 0.5 μl
[0039] Total RNA 1 μl
[0040] Total volume 10 μl
[0041] Reverse transcription reaction...
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