Optimization of ISSR-PCR Reaction System in Pinellia ternata
1 Department of Medicine, Guang'an Vocational & Technical College, Guangan, 638000, China
2 Guang'an Science Park, Guang'an Vocational & Technical College, Guangan, 638000, China
Bioscience Methods, 2023, Vol. 14, No. 2 doi: 10.5376/bm.2023.14.0002
Received: 23 Feb., 2022 Accepted: 20 Apr., 2023 Published: 10 May, 2023
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Yu N., Tang Y.C., Liu Y., and Dai W., 2023, Optimization of ISSR-PCR Reaction System in Pinellia ternata, Bioscience Method, 14(2): 1-8 (doi: 10.5376/bm.2023.14.0002)
In order to establish and perfect ISSR-PCR reaction system of Pinellia ternata, we used P. ternata from Sichuan region as the test material in this research. Genomic DNA of whole plant of P. ternata was used as template, UBC818 primer was used in ISSR-PCR system, L16(45) orthogonal test and single factor experiment were used to optimize the system. Based on the visual analysis of orthogonal experiment, we found the following are the influences of various factors on the experimental results in turn: dNTPs, primer, Mg2+, Taq DNA polymerase, DNA template. Based on single factor screening test, we found the best ISSR-PCR reaction system. The total reaction volume is 20.0 μL, containing dNTPs 0.2 mmol/L, primer 0.8 μmol/L, Mg2+ 2.0 mmol/L, Taq DNA polymerase 0.075 U/μL, DNA 15 ng, annealing temperature 56℃. In this study, ISSR-PCR reaction system was optimized which will be applied to detect the genetic stability of P. ternata tissue culture seedlings.
Pinellia ternata; ISSR-PCR; Optimization