RealPure植物基因組DNA提取試劑盒

● 試劑盒内容：

● 儲存條件(jiàn)和效期：

本試劑盒在常溫（25℃左右）幹燥條件(jiàn)下，可(kě)保存1年(nián)；更長時間的保存可(kě)置于2–8℃。2–8℃保存條件(jiàn)下，若溶液産生(shēng)沉澱，應在使用前置于37℃下溶解沉澱至溶液澄清後再使用。

單獨包裝的RNaseA收到後-20℃保存。

● 産品簡介：

本試劑盒采用可(kě)以特異性結合DNA的離(lí)心吸附柱和獨特的緩沖液系統，能提取多種植物細胞/組織中的基因組DNA。離(lí)心吸附柱可(kě)以高效、專一吸附DNA，可(kě)最大(dà)限度去(qù)除雜質蛋白(bái)及細胞中其他(tā)有機(jī)化合物。提取的基因組DNA片段大(dà)，純度高，質量穩定可(kě)靠。

使用本試劑盒回收的DNA可(kě)适用于各種常規操作(zuò)，包括酶切、PCR、文庫構建、Southern雜交等實驗。

● 提取得(de)率：

● 準備工(gōng)作(zuò)：

1. 準備液氮；65℃水浴；無水乙醇；1.5ml滅菌離(lí)心管。

2. 按照(zhào)标簽所示在緩沖液AP3和漂洗液PW中加入無水乙醇，混勻後蓋緊瓶蓋後常溫貯存備用。

3. 每次使用前請(qǐng)檢查緩沖液AP1，緩沖液AP2和緩沖液AP3是否有沉澱生(shēng)成，如(rú)果出現沉澱，37℃溫浴至沉澱溶解後再使用。

● 操作(zuò)步驟：

如(rú)非指出，所有離(lí)心步驟均爲使用台式離(lí)心機(jī)在常溫下離(lí)心。

1. 取适量植物新鮮組織0.5-1g加入液氮充分(fēn)研磨，取約100mg粉末置于1.5ml離(lí)心管中，加入400 μl 緩沖液AP1和6 μl RNaseA (10mg/ml)，漩渦震蕩1分(fēn)鍾。

2. 将離(lí)心管放(fàng)在65℃水浴10分(fēn)鍾，水浴過程中颠倒離(lí)心管以混合樣品數次。

3. 加入130 μl 緩沖液AP2，颠倒混勻，冰浴5分(fēn)鍾。

4. 12,000rpm離(lí)心5分(fēn)鍾。

注：此步驟離(lí)心去(qù)除去(qù)垢劑，蛋白(bái)以及多糖等雜質。

5. 取上清（通常爲400 μl）轉移到過濾柱CF中，12,000rpm 離(lí)心1分(fēn)鍾。

6. 将濾出液轉移到1.5 ml離(lí)心管中，加入1.5倍體(tǐ)積（例如(rú)400 μl的上清液加600 μl緩沖液AP3）緩沖液AP3（使用前請(qǐng)注意是否已經加入無水乙醇），立即颠倒混勻，簡短(duǎn)離(lí)心以去(qù)除管蓋内壁的水珠。

7．吸取步驟6中的混合液加入到吸附柱CG中（吸附柱放(fàng)入收集管中），12,000 rpm離(lí)心1分(fēn)鍾，倒掉廢液，吸附柱CG放(fàng)回收集管中。

注：離(lí)心吸附柱的最大(dà)容積是700μl，如(rú)果一次不能完全上柱，可(kě)以分(fēn)次上柱離(lí)心，保證全部溶液全部加到離(lí)心柱中。

8．向吸附柱CG中加入500 μl漂洗液PW（使用前請(qǐng)先檢查是否已加入無水乙醇），12,000 rpm離(lí)心1分(fēn)鍾，倒掉廢液，吸附柱CG放(fàng)入收集管中。

9．重複步驟8。

10. 可(kě)選步驟：如(rú)果離(lí)心柱的吸附膜上有顔色，向吸附柱CG中加入500 μl無水乙醇，12,000 rpm離(lí)心1分(fēn)鍾，倒掉廢液，吸附柱CG放(fàng)入收集管中。

11．将吸附柱CG放(fàng)回廢液收集管中，12,000 rpm離(lí)心2分(fēn)鍾。

注：此步驟非常重要，目的是将吸附柱中殘餘的漂洗液去(qù)除。漂洗液中乙醇的殘留會影(yǐng)響後續的酶反應（酶切、PCR等）實驗。

12．将吸附柱CG轉入一個幹淨的1.5ml離(lí)心管中，向吸附膜的中間部位懸空滴加50-100 μl經65℃水浴預熱(rè)的洗脫緩沖液EB，常溫放(fàng)置2分(fēn)鍾，12,000 rpm g離(lí)心2分(fēn)鍾。

注；1. 洗脫緩沖液體(tǐ)積最好不少于50 μl，體(tǐ)積過小影(yǐng)響回收效率。

2. 洗脫液的pH值對于洗脫效率有很大(dà)影(yǐng)響。若用水做洗脫液應保證其pH值在7.0-8.5範圍内，pH值低于7.0會降低洗脫效率。

13.  DNA産物-20℃保存。

● DNA濃度及純度檢測：

基因組DNA片段的大(dà)小與樣品保存時間、操作(zuò)過程中的剪切力等因素有關。提取的DNA片段可(kě)用瓊脂糖凝膠電泳和紫外分(fēn)光(guāng)光(guāng)度計(jì)檢測濃度與純度。可(kě)配制0.8-1.0％瓊脂糖凝膠，使用λ/HindIII判斷基因組的大(dà)小，完整的基因組大(dà)小應在23kb以上。使用分(fēn)光(guāng)光(guāng)度計(jì)檢測時， OD₂₆₀/OD₂₈₀比值應爲1.7–1.9之間，如(rú)果洗脫時不使用洗脫緩沖液，而使用去(qù)離(lí)子水洗脫，比值可(kě)能偏低，但(dàn)并不表明DNA純度不高。

核酸提取産品發表文章(zhāng)

1. [2008 IF=1.749] Development of a sequence-characterized ampli?ed region marker for diagnosis of dwarf bunt of wheat and detection of Tilletia controversa Kuhn.

Author: J.H. Liu, L. Gao, T.G. Liu and W.Q. Chen

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: Letters in Applied Microbiology 2009,49,235-240

Institution：Institute of Plant Protection ,Chinese Academy of Agricultural Sciences

Paper link： https://doi.org/10.1111/j.1472-765X.2009.02645.x

2. [2009 IF=2.435] Characterization of three new S-alleles and development of an S-allele-specific PCR system for rapidly identifying the S-genotype in apple cultivars.

Author: Shenshan Long, Maofu Li, Zhenhai Han, Kun Wang, Tianzhong Li

Product: RTP2201瓊脂糖凝膠回收試劑盒

Journal: Tree Genetics & Genomes (2010) 6:161–168

Institution：China Agricultural University

Paper link： https://link.springer.com/article/10.1007/s11295-009-0237-6

3. [2010 IF=0.921] An ISSR-based Approach for the Molecular Detection and Diagnosis of Dwarf Bunt of Wheat, Caused by Tilletia controversa Kuhn.

Author: Li Gao,Wanquan Chen and Taiguo Liu

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: J Phytopathol 159:155–158 (2011)

Institution：State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, CAAS

Paper link：https://onlinelibrary.wiley.com/doi/10.1111/j.1439-0434.2010.01735.x

4. [2010 IF=1.359] Curing the Plasmid pXO2 from Bacillus anthracis A16 Using Plasmid Incompatibility.

Author: Huagui Wang, Xiankai Liu, Erling Feng, Li Zhu, Dongshu Wang, Xiangru Liao, Hengliang Wang

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: Curr Microbiol (2011) 62:703–709

Institution：State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology

Paper link：https://link.springer.com/article/10.1007/s00284-010-9767-2

5. [2010 IF=1.993] Computation-assisted SiteFinding-PCR for isolating flanking sequence tags in rice

Author: Hongru Wang, Jun Fang, Chengzheng Liang, Minghui He, Qiye Li, and Chengcai Chu

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: BioTechniques Vol. 51 | No. 6 | 2011

Institution：Institute of Genetics and Developmental Biology, Chinese Academy of Sciences

Paper link：https://pubmed.ncbi.nlm.nih.gov/22150334/

6. [2012 IF=0.903] T Polymorphisms in major histocompatibility complex class IIa genes are associated with resistance to infectious hematopoietic necrosis in rainbow trout, Oncorhynchus mykiss (Walbaum, 1792).

Author: Z. Liu, D.-D. Hu, S.-J. Shao, J.-Q. Huang, J.-F. Wang and J. Yang

Product: RTP2202 PCR産物純化試劑盒

Journal: J. Appl. Ichthyol. 29 (2013), 1234–1240

Institution：College of Animal Science and Technology, Gansu Agricultural University

Paper link：https://onlinelibrary.wiley.com/doi/full/10.1111/jai.12326

7. [2012 IF=1.958] Cloning, bioinformatics and the enzyme activity analyses of a phenylalanine ammonia-lyase gene involved in dragon’s blood biosynthesis in Dracaena cambodiana.

Author: Xing-Hong Wang，Changhe Zhang

Product: RTP2202 PCR産物純化試劑盒

Journal: Mol Biol Rep (2013) 40:97–107

Institution：Yunnan Institute of Microbiology, Yunnan University

Paper link：https://link.springer.com/article/10.1007/s11033-012-2032-y

8. [2013 IF=1.687] Tobacco Arabinogalactan Protein NtEPc Can Promote Banana (Musa AAA) Somatic Embryogenesis.

Author: H. Shu & L. Xu & Z. Li & J. Li & Z. Jin & S. Chang

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: Appl Biochem Biotechnol (2014) 174:2818–2826

Institution：Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences

Paper link：https://link.springer.com/article/10.1007/s12010-014-1228-0

9. [2015 IF=1.32] Association between MHC II beta chain gene polymorphisms and resistance to infectious haematopoietic necrosis virus in rainbow trout (Oncorhynchus mykiss, Walbaum, 1792).

Author: Juan Yang, Zhe Liu, Hai-Na Shi, Jiu-Pan Zhang, Jian-Fu Wang, Jin-Qiang Huang & Yu-Jun Kang

Product: RTP2202 PCR産物純化試劑盒

Journal: Aquaculture Research, 2016, 47, 570–578

Institution：College of Animal Science and Technology, Gansu Agricultural University

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10. [2015 IF=] A weird DNA band in PCR and its cause.

Author: Chang Shenghe, Sun Wei, Zhou Zhaoxi, Li Jingyang, Dai Minjie, Shu Haiyan

Product: RTP2102 普通質粒小提試劑盒

Journal: Journal of Plant Science & Molecular Breeding  Volume 5 Article 2 2016

Institution：Haikou experimental station, Chinese Academy of Tropical Agricultural Sciences

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11. [2017 IF=4.076] Application of Real-Time Quantitative PCR to Detect Mink Circovirus in Naturally and Experimentally Infected Minks.

Author: Xingyang Cui, Yunjia Shi, Lili Zhao, Shanshan Gu, Chengwei Wei, Yan Yang,Shanshan Wen, Hongyan Chen and Junwei Ge

Product: RTP2102 普通質粒小提試劑盒

Journal: Fronties in Microbiology May 2018 | Volume 9 | Article 937

Institution：College of Veterinary Medicine, Northeast Agricultural University

Paper link：https://pubmed.ncbi.nlm.nih.gov/29867846

12. [2017 IF=3.974] In vitro and in vivo toxic e?ects and in?ammatory responses induced by carboxylated black carbon-lead complex exposure.

Author: Shuanglin Jiang,, Mengting Shang,Kui Mu，Nan Jiang，Haiyan Wen，Rong Wang，Hai Wu，Wenyong Li

Product: RTG2402 動物/細胞基因組DNA提取試劑盒

Journal: Ecotoxicology and Environmental Safety 165 (2018) 484–494

Institution：Key Laboratory of Embryo Development and Reproductive Regulation of Anhui Province, Fuyang Normal University

Paper link：http://www.sciencedirect.com/science/article/pii/S0147651318309011

13. [2018 IF=8.063] ATF4 destabilizes RET through nonclassical GRP78 inhibition to enhance chemosensitivity to bortezomib in human osteosarcoma

Author: Jie Luo,# Yuanzheng Xia,# Yong Yin, Jun Luo, Mingming Liu, Hao Zhang, Chao Zhang, Yucheng Zhao, Lei Yang, and Lingyi Kong

Product: RTP2101 高純質粒小提試劑盒

Journal: Theranostics 2019, Vol. 9, Issue 21

Institution：School of Traditional Chinese Pharmacy, China Pharmaceutical University

Paper link：https://pubmed.ncbi.nlm.nih.gov/31534554

14. [2020 IF=1.857] Characterization and Developmental Expression Patterns of Four Hexamerin Genes in the Bumble Bee, Bombus terrestris (Hymenoptera: Apidae).

Author: Yakai Tian, Yingping Qu,Kun Dong,Shaoyu He,Wu Jie, and Jiaxing Huang

Product: RTP2201 瓊脂糖凝膠回收試劑盒

Journal: Journal of Insect Science (2021) 21(5): 13; 1–8

Institution：Institute of Apicultural Research, Chinese Academy of Agricultural Sciences

Paper link：https://doi.org/10.1093/jisesa/ieab078

15. [2020 IF=1.336] Isopentenyl Diphosphate Isomerase (IPI) Gene Silencing Negatively Afects Patchouli Alcohol Biosynthesis in Pogostemon cablin

Author: Wuping Yan, Yuzhang Yang, Yougen Wu, Jing Yu, Junfeng Zhang, Dongmei Yang, Zeeshan Ul Haq Muhammad

Product: RTP2102 普通質粒小提試劑盒

Journal: Plant Molecular Biology Reporter Published 27 January 2021

Institution：College of Horticulture, Hainan University

Paper link：https://link.springer.com/article/10.1007/s11105-020-01269-0

16. [2021 IF=6.064] Genome resequencing and transcriptome analysis reveal the molecular mechanism of albinism in Cordyceps militaris.

Author: Ying Zhao, YuDong Liu, Xun Chen and Jun Xiao

Product: RTG2407 真菌基因組DNA提取試劑盒

Journal: Fronties in Microbiology.  Published 11 April 2023

Institution： Institute of Edible Fungi, Liaoning Academy of Agricultural Sciences

Paper link：https://www.frontiersin.org/articles/10.3389/fmicb.2023.1153153/full