A kind of pcr reaction system for scot molecular marker of berry
A molecular marker and reaction system technology, applied in the biological field, to achieve the effects of good repeatability, high sensitivity and simple operation
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Embodiment 1
[0019] The method for establishing the PCR reaction system of the SCoT molecular marker of embodiment 1
[0020] 1 Experimental method
[0021] 1.1 Genomic DNA extraction and detection
[0022] The extraction of plant genomic DNA of berry plant adopts the improved CTAB method (Li Jinlu, Wang Shuo, Yu Jing, Wang Ling, & Zhou Shiliang. (2013). An improved plant DNA extraction method. Acta Botany, 48(1), 72-78.). The DNA concentration and quality were determined by a spectrophotometer, and an appropriate amount of samples were taken for detection by 0.8% agarose electrophoresis, and the rest of the DNA samples were stored in a -20°C refrigerator for later use.
[0023] 1.2 Optimization of SCoT-PCR reaction system
[0024] The reaction system was determined to be 20 μL, and the pre-amplified primer SC2 (5'-CAACAATGGCTACCACCC-3', whose nucleotide sequence is shown in SEQ ID NO.1) was first used as the Adenophyllum genomic DNA amplification system to establish Primary primers, T...
Embodiment 2
[0039] The application example of the PCR reaction system of the SCoT molecular marker of embodiment 2
[0040] On the basis of the optimal SCoT-PCR reaction system, 8 randomly selected samples of A. adenocarpa were amplified using SC35 (5'-CATGGCTACCACCGGCCC-3', whose nucleotide sequence is shown in SEQ ID NO.2) primers .
[0041] By amplifying 8 randomly selected berry samples, the results ( image 3 ) showed that the 8 samples were effectively amplified, and the amplified bands were clear, that is, the established SCoT-PCR reaction system of A. adenifolia was stable.
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