Molecular identification method suitable for amorphophallus konjac and amorphophallus albus hybrid
Through PCR amplification and first-generation DNA sequencing technology based on SNP primers, the instability and reliability of the identification of hybrid subsidiaries in the first generation of flower konjac and white konjac were solved, and efficient and reliable identification of hybrid subsidiaries was achieved.
CN119799950APending Publication Date: 2025-04-11QUJING NORMAL UNIV
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Patent Information
- Application Number
- CN202510041654.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-10
- Publication Date
- 2025-04-11
AI Technical Summary
Technical Problem
The prior art is difficult to effectively identify the hybrid subgeneration of flower konjac and white konjac, and conventional morphological methods are unstable and have poor reliability.
Method used
The molecular identification method based on SNP primers combined with first-generation sequencing technology was used to identify the hybrid subgeneration by PCR amplification and first-generation DNA sequencing through ZJ20 primers, and the hybrid genotype A/T at positions 170-180bp, 190bp-200bp and 210bp-220bp were observed.
Benefits of technology
It has achieved remarkable identification of the hybrid subgeneration of flower konjac and white konjac, which has the advantages of being simple and efficient, reliable results and suitable for large-scale testing.
✦ Generated by Eureka AI based on patent content.
Abstract
The invention belongs to the technical field of plant molecular biology, and discloses a PCR amplification primer suitable for molecular identification of amorphophallus konjac and amorphophallus albus hybrids, the PCR amplification primer comprises a ZJ20 primer, and the sequence of the ZJ20 primer is F: CAGACCATAATCAGCATG; r is GTCTCCACT GCCGTACCTT, and R is GTCTCCACT The molecular identification method comprises the following steps: step 1, extracting genome DNA of a to-be-detected konjak sample, carrying out amplification by using the PCR primer, then carrying out first-generation DNA sequencing, and if a hybrid genotype A / T is observed at three positions of 170-180bp, 190bp-200bp and 210bp-220bp, determining that a hybrid first filial generation of amorphophallus konjac and amorphophallus albus is obtained, and a reaction system for PCR amplification in the step 2 is as follows: the reaction total volume is 25 microliters, and the reaction time is 30 minutes; a reagent contained in the step 3 is as follows: 12.5 mu L of a 2 * PCR Mix premixed solution, 0.3 mu M of R and F primers and 30 ng of a sample genome DNA solution, finally, sterile ultrapure water is used for complementing the deficient volume, and the amplification process of the PCR reaction in the step 3 is as follows: pre-denaturation at 95 DEG C is performed for 5 minutes, then denaturation at 95 DEG C is performed for 45 seconds, annealing at 56 DEG C is performed for 45 seconds, extension at 72 DEG C is performed for 45 seconds, and 35 amplification cycles are performed in total.
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