Optimization of EST-SSR PCR reaction system for <em>Betula luminifera</em>

CHEN Zheng; JIANG Xiao-feng; TONG Zai-kang

doi:10.11833/j.issn.2095-0756.2012.06.023

Volume 29 Issue 6

Nov. 2012

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CHEN Zheng, JIANG Xiao-feng, TONG Zai-kang. Optimization of EST-SSR PCR reaction system for Betula luminifera[J]. Journal of Zhejiang A&F University, 2012, 29(6): 960-965. doi: 10.11833/j.issn.2095-0756.2012.06.023

Citation:

CHEN Zheng, JIANG Xiao-feng, TONG Zai-kang. Optimization of EST-SSR PCR reaction system for Betula luminifera[J]. Journal of Zhejiang A&F University, 2012, 29(6): 960-965. doi: 10.11833/j.issn.2095-0756.2012.06.023

Optimization of EST-SSR PCR reaction system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2012.06.023

1.
The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University Lin’an 311300，Zhejiang，China

More Information

Corresponding author: TONG Zai-kang
Received Date: 2011-10-26
Rev Recd Date: 2012-02-22
Publish Date: 2012-12-20

Abstract

To obtain stable PCR product，providing the basis for further study，orthogonal design was used to optimize an Expressed Sequence Tags （EST）-Simple Sequence Repeat （SSR） Polymerase Chain Reaction （PCR） amplification system for Betula luminifera in terms of five factors containing rTaq DNA polymerase，Mg2+，DNA template，dNTPs and primers in five levels. Results showed an optimal EST-SSR PCR system for B. luminifera was 4 mgL-1 DNA template，0.500 molL-1 primers，1.250 mmolL-1 Mg2+，0.250 mmolL-1 dNTPs，and 0.072 516.67 mkatL-1 rTaq DNA polymerase. And the optimal annealing temperature to each primers for SSR PCR reaction system is determined by gradient PCR. Using the optimal reaction system，stable and clear polymorphic bands were acquired.［Ch，6 fig. 3 tab. 15 ref.］
- forest tree breeding,
- Betula luminifera,
- EST-SSR,
- PCR reaction system,
- Orthogonal design

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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 EST-SSR PCR reaction system for Betula luminifera

1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

Received Date: 2011-10-26

Rev Recd Date: 2012-02-22

Publish Date: 2012-12-20

Key words:

Abstract: To obtain stable PCR product，providing the basis for further study，orthogonal design was used to optimize an Expressed Sequence Tags （EST）-Simple Sequence Repeat （SSR） Polymerase Chain Reaction （PCR） amplification system for Betula luminifera in terms of five factors containing rTaq DNA polymerase，Mg2+，DNA template，dNTPs and primers in five levels. Results showed an optimal EST-SSR PCR system for B. luminifera was 4 mgL-1 DNA template，0.500 molL-1 primers，1.250 mmolL-1 Mg2+，0.250 mmolL-1 dNTPs，and 0.072 516.67 mkatL-1 rTaq DNA polymerase. And the optimal annealing temperature to each primers for SSR PCR reaction system is determined by gradient PCR. Using the optimal reaction system，stable and clear polymorphic bands were acquired.［Ch，6 fig. 3 tab. 15 ref.］

Optimization of EST-SSR PCR reaction system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2012.06.023

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Optimization of EST-SSR PCR reaction system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2012.06.023

1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

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Optimization of EST-SSR PCR reaction system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2012.06.023

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Optimization of EST-SSR PCR reaction system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2012.06.023

1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

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