Amplification primer and amplification method of complete mitochondrial genome sequence of nibea miichthioides chu
A whole genome, amplification primer technology, applied in the field of genetic engineering
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[0042] Using the extracted genomic DNA of yellow aquarium as a template, first amplify the products of 4 large fragments with 4 pairs of specific primers for large fragment amplification, and then use the method of genome walking to use the amplified 4 large fragments respectively. The fragment was used as a template for PCR amplification using 19 primers for genome walking. The PCR product was detected by 1.0% agarose electrophoresis and sent to the biological company for sequencing. The sequencing results were assembled using biological software. The specific steps are as follows:
[0043] 1. 5×TBE Buffer preparation
[0044] Weigh 54g of tromethamine, Na 2 EDTA·2H 2 O 3.72g, 27.5g boric acid in a 1L beaker; add about 800ml deionized water and stir evenly; adjust the pH to 8.3 with NaOH, add deionized water to a constant volume to 1L, and store at room temperature for later use.
[0045] 2. DNA extraction
[0046] The high-salt method is used to extract DNA from koala croaker, the s...
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