An SSR molecular labeling technique system for <em>Betula luminifera</em>

YOU Wei-yan; HUANG Hua-hong; CHENG Long-jun; TONG Zai-kang; ZHU Yu-qiu

doi:10.11833/j.issn.2095-0756.2010.03.023

Volume 27 Issue 3

Jun. 2010

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YOU Wei-yan, HUANG Hua-hong, CHENG Long-jun, TONG Zai-kang, ZHU Yu-qiu. An SSR molecular labeling technique system for Betula luminifera[J]. Journal of Zhejiang A&F University, 2010, 27(3): 464-469. doi: 10.11833/j.issn.2095-0756.2010.03.023

Citation:

YOU Wei-yan, HUANG Hua-hong, CHENG Long-jun, TONG Zai-kang, ZHU Yu-qiu. An SSR molecular labeling technique system for Betula luminifera[J]. Journal of Zhejiang A&F University, 2010, 27(3): 464-469. doi: 10.11833/j.issn.2095-0756.2010.03.023

An SSR molecular labeling technique system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2010.03.023

1.
School of Forestry and Biotechnology，Zhejiang Forestry College，Lin’an 311300，Zhejiang，China

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Corresponding author: TONG Zai-kang
Publish Date: 2010-06-20

Abstract

To find the optimal simple sequence repeats（SSR） system for Betula luminifera，the cetyltrimethyl-ammonium bromide （CTAB） method was used to extract DNA from tender leaves，using the known primers from related species：B. platyphylla var. japonica and B. pendula. Then，one factor was changed at a time to optimize the polymerase chain reaction（PCR）. Next，Polyacrylamide Gel Electrophoresis（PAGE） was used to detect the amplified products with three primers segments randomly selected from thirty-six pairs of primers and sequenced to further check the universality of the primer. Results showed that in a volume of 20 L，the optimal reaction system was 60 ng template DNA，1.500 mmolL－1 Mg2+，0.175 mmolL－1 deoxyribonucleotide triphosphate（dNTP）s，2.500 mmolL－1 primer，and 1.0 16.67 nkat Taq polymerase. Compared with B. platyphylla var. japonica and B. pendula，the optimal annealing temperature of the primer was 1C higher. The blast results from primer AF310851，AB084479，and AB084480 showed that the values of max ident were all above 95.0%. This indicated that the SSR primers of birch could be used with B. luminifera.［Ch，7 fig. 2 tab. 17 ref.］
- forest tree breeding,
- Betula luminifera,
- simple sequence repeats （SSR） marker

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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An SSR molecular labeling technique system for Betula luminifera

1. School of Forestry and Biotechnology，Zhejiang Forestry College，Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

Publish Date: 2010-06-20

Key words:

Abstract: To find the optimal simple sequence repeats（SSR） system for Betula luminifera，the cetyltrimethyl-ammonium bromide （CTAB） method was used to extract DNA from tender leaves，using the known primers from related species：B. platyphylla var. japonica and B. pendula. Then，one factor was changed at a time to optimize the polymerase chain reaction（PCR）. Next，Polyacrylamide Gel Electrophoresis（PAGE） was used to detect the amplified products with three primers segments randomly selected from thirty-six pairs of primers and sequenced to further check the universality of the primer. Results showed that in a volume of 20 L，the optimal reaction system was 60 ng template DNA，1.500 mmolL－1 Mg2+，0.175 mmolL－1 deoxyribonucleotide triphosphate（dNTP）s，2.500 mmolL－1 primer，and 1.0 16.67 nkat Taq polymerase. Compared with B. platyphylla var. japonica and B. pendula，the optimal annealing temperature of the primer was 1C higher. The blast results from primer AF310851，AB084479，and AB084480 showed that the values of max ident were all above 95.0%. This indicated that the SSR primers of birch could be used with B. luminifera.［Ch，7 fig. 2 tab. 17 ref.］

An SSR molecular labeling technique system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2010.03.023

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An SSR molecular labeling technique system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2010.03.023

1. School of Forestry and Biotechnology，Zhejiang Forestry College，Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

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An SSR molecular labeling technique system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2010.03.023

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An SSR molecular labeling technique system for Betula luminifera

doi: 10.11833/j.issn.2095-0756.2010.03.023

1. School of Forestry and Biotechnology，Zhejiang Forestry College，Lin’an 311300，Zhejiang，China

Corresponding author: TONG Zai-kang

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