|Table of Contents|

Optimization of Trolliuschinensis Bunge ISSR-PCR Reaction System and Primer Screening

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2025年17
Page:
10-18
Research Field:
Publishing date:

Info

Title:
Optimization of Trolliuschinensis Bunge ISSR-PCR Reaction System and Primer Screening
Author(s):
DONG Menghan; MA Menglu; WANG Ruikang; ZHANG Qin
(College of Landscape Architecture and Tourism,Hebei Agricultural University,Baoding,Hebei 071000)
Keywords:
Trollius chinensis Bunge; ISSR-PCR; primer screening; reaction system
PACS:
S 326
DOI:
10.11937/bfyy.20250074
Abstract:
Taking Trollius chinensis Bunge as experimental material，the optimal reaction system was combined by single factor horizontal screening and orthogonal experimental methods.The effects of four key factors on the system were systematically analyzed，included DNA concentration，Mix enzyme dosage，primer concentration，reaction cycle times，and the primers and their annealing temperature were determined.The aim was to establish the ISSR-PCR reaction system of Trollius chinensis，in order to provide reference for germplasm innovation and genetic diversity analysis of Trollius chinensis Bunge.The results showed that the influence of four factors on the reaction system was PCR cycles>DNA concentration>amount of Mix enzyme used>primer concentration.The best reaction system was,DNA concentration 35 ng·μL-1，primer concentration 9 μmol·L-1，and Mix enzyme amount 9 μL.Reaction procedure,94 ℃ pre-denaturation for 5 minutes，94 ℃ denaturation for 30 seconds，annealing time for 45 seconds，72 ℃ extension for 45 seconds，33 cycles，72 ℃ final extension for 7 minutes，4 ℃ preservation.The best reaction system of ISSR molecular markers was established and 26 ISSR primers were selected，which could be applied to the subsequent germplasm resources research work.

References:

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Last Update: 2025-09-29