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

Turn off MathJax

Article Contents

Article Navigation > Journal of Zhejiang A&F University > 2006 > 23(5): 516-520

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

Relative Article

[1]	HE Ziqiang, ZHANG Huiling, YANG Zhengxiong. Extraction of polysaccharides of Abelmoschus esculentus using ultrasonic-assisted [Bmim] Cl-K₂HPO₄ aqueous two phases optimized by response surface method . Journal of Zhejiang A&F University, 2020, 37(3): 547-555. doi: 10.11833/j.issn.2095-0756.20190339
[2]	ZHANG Jie, YIN Dejie, GUAN Haiyan, QU Qiqi, DONG Li. An overview of Sedum spp. Research . Journal of Zhejiang A&F University, 2018, 35(6): 1166-1176. doi: 10.11833/j.issn.2095-0756.2018.06.022
[3]	CUI Jing, WU Jigui, HUANG Bogao, JIANG Wanjie, FAN Yaqian, CHENG Jin. The mechanisms of reproductive isolation in orchids . Journal of Zhejiang A&F University, 2016, 33(4): 695-702. doi: 10.11833/j.issn.2095-0756.2016.04.020
[4]	TIAN Fengguang, HE Yingfei, DUAN Chengli. An ISSR-PCR reaction system for Paris polyphylla . Journal of Zhejiang A&F University, 2014, 31(6): 983-989. doi: 10.11833/j.issn.2095-0756.2014.06.023
[5]	CHEN Fang, ZHANG Guangqun, CUI Kunpeng, WANG Hangjun. Design and implementation of an embedded automatic plant recognition system . Journal of Zhejiang A&F University, 2013, 30(3): 379-384. doi: 10.11833/j.issn.2095-0756.2013.03.012
[6]	WU Chong, ZHONG Chonglu, ZHANG Yong, ＪＩＡＮＧ Qingbin, CHEN Yu, CHEN Zhen. Establishment and optimization of an ISSR-PCR system for Chukrasia tabularis . Journal of Zhejiang A&F University, 2013, 30(4): 620-626. doi: 10.11833/j.issn.2095-0756.2013.04.024
[7]	WU Yueyan, TAO Qiaojing, LI Bo, XU Danye. An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker . Journal of Zhejiang A&F University, 2013, 30(6): 844-851. doi: 10.11833/j.issn.2095-0756.2013.06.007
[8]	JIANG Xiaofeng, GAO Yanhui, TONG Zaikang, HUANG Chunhong. Establishing and optimizing a SCoT-PCR system for Lycoris . Journal of Zhejiang A&F University, 2013, 30(3): 444-452. doi: 10.11833/j.issn.2095-0756.2013.03.023
[9]	JIANG Xiao-ling, HUANG Qiu-xian, LI Hong, ZHAO Jia-ping. A review of advances in plant thaumatin-like proteins . Journal of Zhejiang A&F University, 2012, 29(2): 279-287. doi: 10.11833/j.issn.2095-0756.2012.02.019
[10]	CHEN Zheng, JIANG Xiao-feng, TONG Zai-kang. Optimization of EST-SSR PCR reaction system for Betula luminifera . Journal of Zhejiang A&F University, 2012, 29(6): 960-965. doi: 10.11833/j.issn.2095-0756.2012.06.023
[11]	GAO Hang-yang, ZHANG Qi-xiang, HU Heng-kang, ZHANG Zhen-ru, HUANG Jian-qin. Optimization of a rooting culture system for Rhododendron fortunei . Journal of Zhejiang A&F University, 2011, 28(6): 982-985. doi: 10.11833/j.issn.2095-0756.2011.06.024
[12]	YE Sheng-yue, CHEN Gang, ZHANG Hui, LIU Jian-jie, ZENG Yan-ru, WU Hui-min. Optimization of an ISSR-PCR system and primer screening for Torreya grandis ‘Merrillii’ . Journal of Zhejiang A&F University, 2010, 27(1): 87-92. doi: 10.11833/j.issn.2095-0756.2010.01.014
[13]	GU Cui-hua, WANG Shou-xian, ZHANG Qi-xiang. Optimization of AFLP molecular marker system in Lagerstroemia plants in China . Journal of Zhejiang A&F University, 2008, 25(3): 298-303.
[14]	CAO Fu-liang, WANG Guo-xia, LI Guang-ping, HUA Zhe-bin. Optimizing the inter simple sequence repeat(ISSR) reaction system with Gingko biloba . Journal of Zhejiang A&F University, 2008, 25(2): 186-190.
[15]	YANG Jing-yuan, XIONG Dan, LIANG Hong-wei, WANG Yu-bing, LIAOMing-ya, CHEN Fa-ju. Optimal reaction system for random amplified polymorphic DNA with endangered Emmenopterys henryi . Journal of Zhejiang A&F University, 2007, 24(3): 279-283.
[16]	ZHANG Lei-fan, GAO Yan-hui, ZHUYu-qiu, LIUZhi-gao, TONG Zai-kang, HUANG Hua-hong. An inter simple sequence repeats(ISSR)reaction system for Lycoris(Amaryllidaceae) . Journal of Zhejiang A&F University, 2007, 24(2): 156-161.
[17]	YANG Ping, ZHENG Qian-jian, ZHAO Ming-shui, TANG Ding-qin. Optimization of the reaction system for CAPS marker analysis in Cryptomeria fortunei . Journal of Zhejiang A&F University, 2006, 23(1): 52-55.
[18]	LIN Xin-chun, YU Zhi-xiong. Characters of leaf epidermis of Magnoliaceae and its taxonomic significance . Journal of Zhejiang A&F University, 2004, 21(1): 33-39.
[19]	WANG Guo-liang. On restoration ecology of Castanopsis eyrei forest in Mount Longxi of Fujian (Ⅰ) . Journal of Zhejiang A&F University, 2002, 19(4): 363-366.
[20]	BI Li-jun. Extracting flavonoid compounds from Toona sinensis leaves . Journal of Zhejiang A&F University, 2000, 17(2): 146-149.

References
Proportional views

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

Get Citation

PDF

XML

Article views(1403) PDF downloads(254) Cited by()

Proportional views

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

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Related

Proportional views

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

HTML

Catalog

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

Abstract

Relative Article

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Related

Proportional views

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

HTML

Catalog

Export File

Citation

Format

Content

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