Potato 1-amino-cyclopropane-1-carboxylic-synthesized zyme gene chip and primer
A technology of aminocyclopropane and gene fragments, applied in the field of nucleic acids
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[0018] Synthesis of ACC primers
[0019] Primers were prepared according to conventional methods well known in the art. For example, the synthesis is carried out on an automatic DNA synthesizer (such as ABI3949 automatic DNA synthesizer from PE Company, USA) according to the manufacturer's instructions. The forward primer 5'-gctgaggagtgctgggatt-3'(SEQ ID NO:2) of potato ACC synthase gene and the reverse primer 5'-ccttgtttccatgaaatggtg-3'(SEQ ID NO:3) of potato ACC synthase gene were synthesized respectively.
[0020] Amplification of 265bp DNA fragment of ACC gene
[0021] Potato DNA was extracted with the Qiagen DNeasy Plant Mini kit according to the manufacturer's instructions and dissolved in 100 microliters of double distilled water. Take 5 μl of DNA and add PCR master mix (1x PCR buffer, 200 μmol dNTPs, 200 μmol potato ACC forward and reverse primers and 0.25 units Taq DNA polymerase). The PCR buffer is: 10mM Tris-HCl (pH8.6), 50mM KCl, 1.5mM MgCl 2 and 0.1% Triton X-...
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