Establishment and optimization of an ISSR-PCR system for<em> Chukrasia tabularis</em>

WU Chong; ZHONG Chonglu; ZHANG Yong; ＪＩＡＮＧ Qingbin; CHEN Yu; CHEN Zhen

doi:10.11833/j.issn.2095-0756.2013.04.024

Volume 30 Issue 4

Jul. 2013

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WU Chong, ZHONG Chonglu, ZHANG Yong, ＪＩＡＮＧ Qingbin, CHEN Yu, CHEN Zhen. Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis[J]. Journal of Zhejiang A&F University, 2013, 30(4): 620-626. doi: 10.11833/j.issn.2095-0756.2013.04.024

Citation:

WU Chong, ZHONG Chonglu, ZHANG Yong, ＪＩＡＮＧ Qingbin, CHEN Yu, CHEN Zhen. Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis[J]. Journal of Zhejiang A&F University, 2013, 30(4): 620-626. doi: 10.11833/j.issn.2095-0756.2013.04.024

Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

doi: 10.11833/j.issn.2095-0756.2013.04.024

1.
Research Institute of Tropical Forestry，Chinese Academy of Forestry，Guangzhou 510520，Guangdong，China

More Information

Corresponding author: ZHONG Chonglu
Received Date: 2012-06-19
Rev Recd Date: 2012-10-04
Publish Date: 2013-08-20

Abstract

The genomic DNA from twenty-four varieties of germplasm material for Chukrasia tabularis leaves was systematically analyzed for template DNA，primer，dNTPs，Mg2+ concentration，dosage of Taq DNA polymerase，and annealing temperature with an inter simple sequence repeat（ISSR）-polymerase chain reaction（PCR）. Results showed that the optimal reaction system for the ISSR was a 20 L system containing 30 ng template DNA，1.00 molL－1 random primers, 0.15 mmolL－1 dNTPs，2.50 mmolL－1 Mg2+， and 2.50 16.67 nkat Taq DNA polymerase. The optimal annealing temperature was 56 ℃ with the PCR procedure pre-denaturized at 94 ℃ for 5.0 min, denaturized at 94 ℃ for 45 s, and annealed at 56 ℃ for 45 s with an extension at 72 ℃ for 1.5 min，a reaction of 40 cycles，and re-extension at 72 ℃ for 7 min. These products were then stored at 4 ℃. Results indicated that stable bands could be amplified. ［Ch，8 fig. 1 tab. 27 ref.］
- forest tree breeding,
- Chukrasia tabularis,
- inter simple sequence repeat （ISSR）,
- polymerase chain reaction （PCR） conditions,
- optimization

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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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Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

1. Research Institute of Tropical Forestry，Chinese Academy of Forestry，Guangzhou 510520，Guangdong，China

Corresponding author: ZHONG Chonglu

Received Date: 2012-06-19

Rev Recd Date: 2012-10-04

Publish Date: 2013-08-20

Key words:

forest tree breeding /
Chukrasia tabularis /
inter simple sequence repeat （ISSR） /
polymerase chain reaction （PCR） conditions /
optimization

Abstract: The genomic DNA from twenty-four varieties of germplasm material for Chukrasia tabularis leaves was systematically analyzed for template DNA，primer，dNTPs，Mg2+ concentration，dosage of Taq DNA polymerase，and annealing temperature with an inter simple sequence repeat（ISSR）-polymerase chain reaction（PCR）. Results showed that the optimal reaction system for the ISSR was a 20 L system containing 30 ng template DNA，1.00 molL－1 random primers, 0.15 mmolL－1 dNTPs，2.50 mmolL－1 Mg2+， and 2.50 16.67 nkat Taq DNA polymerase. The optimal annealing temperature was 56 ℃ with the PCR procedure pre-denaturized at 94 ℃ for 5.0 min, denaturized at 94 ℃ for 45 s, and annealed at 56 ℃ for 45 s with an extension at 72 ℃ for 1.5 min，a reaction of 40 cycles，and re-extension at 72 ℃ for 7 min. These products were then stored at 4 ℃. Results indicated that stable bands could be amplified. ［Ch，8 fig. 1 tab. 27 ref.］

Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

doi: 10.11833/j.issn.2095-0756.2013.04.024

Abstract

References

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Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

doi: 10.11833/j.issn.2095-0756.2013.04.024

1. Research Institute of Tropical Forestry，Chinese Academy of Forestry，Guangzhou 510520，Guangdong，China

Corresponding author: ZHONG Chonglu

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Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

doi: 10.11833/j.issn.2095-0756.2013.04.024

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Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis

doi: 10.11833/j.issn.2095-0756.2013.04.024

1. Research Institute of Tropical Forestry，Chinese Academy of Forestry，Guangzhou 510520，Guangdong，China

Corresponding author: ZHONG Chonglu

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