|Table of Contents|

Establishment and Optimalization of ISSR-PCR Reaction System on Rhododendron molle

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2020年21
Page:
96-102
Research Field:
Publishing date:

Info

Title:
Establishment and Optimalization of ISSR-PCR Reaction System on Rhododendron molle
Author(s):
QIN Fang1; SHI Yancai2; ZOU Rong2; TANG Jianmin2; JIANG Yunsheng2; XIONG Zhongchen2
(1.College of Science,Guangxi Normal University,Guilin,Guangxi 541006;2.Guangxi Institute of Botany,Guangxi Zhuangzu Autonomous Region and the Chinese Academy of Sciences,Guilin,Guangxi 541006)
Keywords:
Rhododendron molle; ISSR-PCR reaction system; orthogonal test; single-factor experiment
PACS:
-
DOI:
10.11937/bfyy.20194897
Abstract:
The leaves of Rhododendron molle were used as the test materials.Through orthogonal design and single factor test,to establish ISSR-PCR program for Rhododendron molle.Five factors were tested by orthogonal design and single factor test,Mg2+ concentration,dNTP concentration,Taq DNA polymerase concentration,primer concentration,template DNA concentration and reaction procedure.The results showed that,the Taq DNA polymerase amount was 2.5 U,the primer concentration was 0.2 μmol·L-1,the Mg2+ concentration was 1.0 mmol·L-1,the dNTPs concentration was 0.4 mmol·L-1,and the DNA concentration was 60 ng.Amplification procedure:pre-denaturation for 5 minutes at 94 ℃;denaturation for 30 seconds at 94 ℃,annealing for 30 seconds at 49.6 ℃,extension for 30 seconds at 72 ℃,45 cycles of the above three steps,and extension for 5 minutes at 72 ℃.The establishment of this optimization system could be helpful for further research on the genetic diversity of germplasm resources of Rhododendron molle.

References:

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Last Update: 2021-01-29