Optimization of ISSR-PCR reaction system of <em>Castanopsis eyrei</em> by orthogonal design

ZHANG Wen-biao; JIN Ze-xin; LI Jun-min; PAN Guan-qiong

Volume 23 Issue 5

Feb. 2017

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ZHANG Wen-biao, JIN Ze-xin, LI Jun-min, PAN Guan-qiong. Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design[J]. Journal of Zhejiang A&F University, 2006, 23(5): 516-520.

Citation:

ZHANG Wen-biao, JIN Ze-xin, LI Jun-min, PAN Guan-qiong. Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design[J]. Journal of Zhejiang A&F University, 2006, 23(5): 516-520.

Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

1.
Key Laboratory of the Three Gorge Reservoir Regions Eco-Environment of Ministry of Education, Southwest China Normal University , Chongqing 400715 , China;
2.
Institute of Ecology , Taizhou University , Linhai 317000 , Zhejiang , China

Funds:

浙江省自然科学基金资助项目(Y504220)

Received Date: 2005-10-31
Rev Recd Date: 2006-05-15
Publish Date: 2006-10-20

Abstract

Obtaining the stable and repeatable ISSR-PCR amplification result is the basic work to study genetic diversity of Castanopsis eyrei .The ISSR-PCR amplification system was optimized by using orthogonal design in six factors and five levels (Mg2 + , dNTP , primer , Taq DNA polymerase , bovine serum albumin and template DNA).The suitable reaction conditions of ISSR-PCR were shown as follows :1 Taq buffer (10 mmolL-1 Tris-HCl , pH 9.0 , 50 mmolL-1 KCl , 10 gL-1 TritonX-100), 1.7 mmolL-1 Mg 2+ , 0.25 mmolL-1 dNTP , 8.34 nkat Taq DNA polymerase , 1.5 gL-1 bovine serum albumin , 6 pmol primer , 12 ng template DNA in 10 L reaction volume .The optimal annealing temperature for primer UBC 846 was 56.3 ℃.[ Ch , 2 fig .1 tab .23 ref .]
- botany,
- ISSR-PCR,
- orthogonal design,
- optimization,
- Castanopsis eyrei

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

1. Key Laboratory of the Three Gorge Reservoir Regions Eco-Environment of Ministry of Education, Southwest China Normal University , Chongqing 400715 , China;

2. Institute of Ecology , Taizhou University , Linhai 317000 , Zhejiang , China

Funds:

浙江省自然科学基金资助项目(Y504220)

Received Date: 2005-10-31

Rev Recd Date: 2006-05-15

Publish Date: 2006-10-20

Key words:

Abstract: Obtaining the stable and repeatable ISSR-PCR amplification result is the basic work to study genetic diversity of Castanopsis eyrei .The ISSR-PCR amplification system was optimized by using orthogonal design in six factors and five levels (Mg2 + , dNTP , primer , Taq DNA polymerase , bovine serum albumin and template DNA).The suitable reaction conditions of ISSR-PCR were shown as follows :1 Taq buffer (10 mmolL-1 Tris-HCl , pH 9.0 , 50 mmolL-1 KCl , 10 gL-1 TritonX-100), 1.7 mmolL-1 Mg 2+ , 0.25 mmolL-1 dNTP , 8.34 nkat Taq DNA polymerase , 1.5 gL-1 bovine serum albumin , 6 pmol primer , 12 ng template DNA in 10 L reaction volume .The optimal annealing temperature for primer UBC 846 was 56.3 ℃.[ Ch , 2 fig .1 tab .23 ref .]

Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

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Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

1. Key Laboratory of the Three Gorge Reservoir Regions Eco-Environment of Ministry of Education, Southwest China Normal University , Chongqing 400715 , China;

2. Institute of Ecology , Taizhou University , Linhai 317000 , Zhejiang , China

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Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

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Optimization of ISSR-PCR reaction system of Castanopsis eyrei by orthogonal design

1. Key Laboratory of the Three Gorge Reservoir Regions Eco-Environment of Ministry of Education, Southwest China Normal University , Chongqing 400715 , China; 2. Institute of Ecology , Taizhou University , Linhai 317000 , Zhejiang , China

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1. Key Laboratory of the Three Gorge Reservoir Regions Eco-Environment of Ministry of Education, Southwest China Normal University , Chongqing 400715 , China;

2. Institute of Ecology , Taizhou University , Linhai 317000 , Zhejiang , China