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精準定量 DNA Marker I plus
● 產(chǎn)品編號及規(guī)格:
RTM436-02 100T(2×250μl)
本產(chǎn)品是由7條帶狀雙鏈DNA條帶組成,適用于瓊脂糖凝膠電泳中DNA條帶的分析。產(chǎn)品中已含有1×loading buffer,可根據(jù)實驗需要,直接取2-5 μl電泳,使用方便,電泳圖像清晰。7條帶的大小和含量分別為100(50 ng/5 μl),200(50 ng/μl),300(50 ng/μl),400(100 ng/μl),500(50 ng/μl),600(50 ng/μl),700(50 ng/μl)bp。其中400bp條帶加亮顯示。
● 儲存及運輸:
4℃貯存3個月, -20℃長期貯存;常溫運輸。
● 使用方法:
1. 取2-5 μl本產(chǎn)品加入到瓊脂糖凝膠的加樣孔中就行電泳。
注:根據(jù)電泳梳子的厚度和寬度確定上樣量。每1mm×1mm(厚度×寬度)加樣孔上樣1μl;窄齒梳子(一般為1mm×2mm)上樣2μl;寬齒梳子(一般為1mm×5mm)上樣5μl。如果使用厚齒梳子或?qū)捰?/span>5mm的梳子,可以適當調(diào)整上樣量。
2. 建議電泳條件:凝膠濃度為2.0%,凝膠長度5-7 cm,電泳電壓~7 v/cm,電泳時間30-35分鐘。
3. 通過核酸染料染色后紫外燈或藍光儀下觀察條帶。
● 注意事項:
1. 瓊脂糖的質(zhì)量對DNA的電泳有很大影響,電泳時請盡量使用質(zhì)量優(yōu)等的瓊脂糖。由于PAGE電泳的特殊性,該產(chǎn)品不建議用于PAGE電泳。
2. 使用與電泳緩沖液一致的緩沖液融化瓊脂糖,例如使用1×TAE緩沖液電泳,需使用1×TAE緩沖液溶膠。
3. 多次電泳后緩沖液電導增強,相同電壓下電流增大,導致分辨率降低,并且明顯產(chǎn)熱,嚴重時會引起凝膠熔化或DNA變性。
4. 推薦使用RealSafe系列染料(貨號:GR002,GR003,GR004)染色。如果使用Goldview或EB等染料進行染色,由于此類染料帶更多的正電荷(染料移動方向與核酸泳動方向相反),隨著電泳時間延長,染料向大片段聚集,凝膠會出現(xiàn)陰陽膠現(xiàn)象(染料含量高的凝膠紫外下較亮,含量低的凝膠紫外下發(fā)暗),導致小分子量DNA顯色很弱或不可見。陰陽膠可以在電泳結(jié)束后浸泡染色,將膠浸沒在含1 μg /ml EB或Goldview的電泳緩沖液中20-25 min,小分子量DNA會顯色清楚。更長時間浸泡會因DNA分子擴散而使條帶彌散,小分子量DNA更易彌散。
5. 紫外照射下EB易分解,使DNA條帶熒光變淺。DNA受紫外光照射形成嘧啶二聚體,不利于待回收片段的下游工作,因此盡可能減少紫外照射時間。
6. 蔗糖會結(jié)合DNA,降低其電泳遷移率,建議DNA樣品上樣使用含甘油的Loading Buffer。
7. 大部分核酸工具酶會結(jié)合DNA,降低其電泳遷移率,大分子量DNA尤為明顯。
● 實驗示例:
● 附錄:
DNA在瓊脂糖凝膠中的有效分離范圍
瓊脂糖濃度%(w/v) |
雙鏈DNA有效分離范圍(bp) |
優(yōu)選電泳緩沖液 |
示蹤染料相當于雙鏈DNA片段粗略大小(bp) |
|||
溴酚藍 |
二甲苯菁 |
|||||
TBE |
TAE |
TBE |
TAE |
|||
0.5 |
2000-50000 |
1×TAE |
750 |
1150 |
13000 |
16700 |
0.8 |
800-10000 |
1×TAE |
320 |
530 |
4830 |
6500 |
1.0 |
400-8000 |
1×TAE |
220 |
370 |
3030 |
4160 |
1.2 |
300-7000 |
1×TAE/0.5×TBE |
160 |
275 |
2070 |
2890 |
1.5 |
200-3000 |
1×TAE/0.5×TBE |
110 |
190 |
1300 |
1840 |
2.0 |
100-2000 |
0.5×TBE |
65 |
120 |
710 |
1040 |
3.0 |
25-1000 |
0.5×TBE |
30 |
60 |
300 |
460 |
使用DNA Marker產(chǎn)品發(fā)表部分文章列表:
1. [2021 IF=1.17] Dendrostoma covidicola sp. nov. (Erythrogloeaceae, Diaporthales) on Fagus sylvatica from Sichuan Province, China.
Author: MILAN C. SAMARAKOON, ITTHAYAKORN PROMPUTTHA & JIAN-KUI (JACK) LI
Marker: RTM415,D2000 DNA ladder
Journal: Phytotaxa. 2021,483 (2): 085–094.
Institution:University of Electronic Science and Technology of China
Paper link:https://doi.org/10.11646/phytotaxa.483.2.1
2.[2021 IF=6.057] MUC1 detection and in situ imaging method based on aptamer conformational switch and hybridization chain reaction.
Marker :RTM403,DNA Marker III
Author: ZihanSong,ShengCai.
Journal: Talanta. Vol 239, 2022, 123129
Institution: Zhejiang University
Paper link:https://doi.org/10.1016/j.ijbiomac.2021.11.194
3. [2021 IF=3.24] Curing of Plasmid pXO1 from Bacillus anthracis Using Plasmid Incompatibility.
Marker :RTM404,DNA Marker IV
Author: Xiankai Liu, Dongshu Wang, Huagui Wang, Erling Feng, Li Zhu, Hengliang Wang
Journal: PLos one,January 2012 | Volume 7 | Issue 1 | e29875
Institution:State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology
Paper link: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0029875
4. [2021 IF=3.24] Identification of a Novel Fungus, Leptosphaerulina chartarum SJTU59 and Characterization of Its Xylanolytic Enzymes.
Marker :RTM409,100bp DNA ladder
Author: Qiong Wu, Yaqian Li, Yingying Li, Shigang Gao, Meng Wang, Tailong Zhang, Jie Chen
Journal: PLos one,September 2013 | Volume 8 | Issue 9 | e737
Institution:School of Agriculture and Biology, Shanghai Jiao Tong University
Paper link:https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0073729
5. [2021 IF=3.08] Low genetic diversity and local adaptive divergence of Dracaena cambodiana (Liliaceae) populations associated with historical population bottlenecks and natural selection: an endangered long-lived tree endemic to Hainan Island, China.
Marker :RTM409,100bp DNA ladder
Author: D.-J. Zheng, L.-S. Xie, J.-H. Zhu,Z.-L. Zhang
Journal: Plant Biology,14 (2012) 828–838
Institution:Key Laboratory of Crop Genetics and Breeding of Hainan Province, Hainan Academy of Agricultural Sciences
Paper link:https://onlinelibrary.wiley.com/doi/10.1111/j.1438-8677.2011.00559.x
6. [2021 IF=3.24] Envelope Proteins ofWhite Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT).
Marker :RTM415,D2000 DNA ladder
Author: Shijun Xie, Xiaojun Zhang, Jiquan Zhang, Fuhua Li, Jianhai Xiang
Journal: Plos One,Published: December 21, 2015
Institution:Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Paper link: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0144922
7. [2021 IF=1.17] Rhopalostroma brevistipitatum sp. nov. from Thailand with an extended generic description for Rhopalostroma.
Marker :RTM415,D2000 DNA ladder
Author: DINUSHANI A. DARANAGAMA, XINGZHONG LIU, SUNITA CHAMYUANG, MARC STADLER,ALI H.BAHKALI, KEVIN D. HYDE
Journal: Phytotaxa,2015,227 (3): 229–242
Institution:State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences
Paper link:https://www.biotaxa.org/Phytotaxa/article/view/phytotaxa.227.3.2/0
8. [2021 IF=20.37] A multi-locus phylogenetic evaluation of Diaporthe (Phomopsis)
Marker :RTM415,D2000 DNA ladder
Author: Dhanushka Udayanga & Xingzhong Liu &Pedro W. Crous & Eric H. C. McKenzie &Ekachai Chukeatirote & Kevin D. Hyde
Journal: Fungal Diversity (2012) 56:157–171
Institution:State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences
Paper link:https://link.springer.com/article/10.1007/s13225-012-0190-9
9. [2021 IF=2.9] An easily-performed high-throughput method for plant genomic DNA extraction.
Marker :RTM421,λ DNA/HindIII
Author: Deshui Yu, Ju Zhang, Guangxuan Tan, Ningshu Yu, Qiuyue Wang, Qiqi Duan, Xin
Qi, Mingjiao Cheng, Chunxue Yan, Zhangkun Wei, Zhenmiao Yu, Wenchao Huang, Chengwei Li
Journal: Analytical Biochemistry Volume 569, 15 March 2019, Pages 28-30
Institution:Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal
University
Paper link:
https://www.sciencedirect.com/science/article/abs/pii/S0003269718310637?via%3Dihub
10. [2021 IF=2.88] Genetic diversity and genetic structure of Decalobanthus boisianus in Hainan Island, China.
Marker :RTM415,D2000 DNA ladder
Author: Huan Jiang,Wenxing Long, Hui Zhang,Chengneng Mi,Tao Zhou,Zongzhu Chen
Journal: Ecology and Evolotion Volume 569, 15 March 2019, Pages 28-30
Institution:Institute of Tropical Agriculture and Forestry, Hainan University
Paper link: https://doi.org/10.1002/ece3.5127
11. [2021 IF=5.64] Identi?cation of a Novel Hypovirulence-Inducing Hypovirus From Alternaria alternata.
Marker :RTM406,DNA Marker VI
Author: Huan Li, Ruiling Bian, Qian Liu, Liu Yang, Tianxing Pang, Lakha Salaipeth,
Ida Bagus Andika, Hideki Kondo and Liying Sun
Journal: Frontiers in Microbiology May 2019 | Volume 10 | Article 1076
Institution:State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University
Paper link: https://www.frontiersin.org/articles/10.3389/fmicb.2019.01076/full
12. [2021 IF=2.6] Natural occurrence of Alternaria mycotoxins in wheat and potential of reducing associated risks using magnolol.
Marker :RTM415,D2000 DNA ladder
Author: Dongmei Jiang, Dizhe Wei, Hongtao Li, Liuqing Wang, Nan Jiang,Yunchao Li and Meng Wang
Journal: Journal of the Science of Food and Agriculture Volume101, Issue7,2021,3071-3077
Institution:Beijing Research Center for Agricultural Standards and Testing, Beijing Academy of Agriculture and Forestry Sciences
Paper link:https://onlinelibrary.wiley.com/doi/10.1002/jsfa.10901
13. [2021 IF=20.37] Taxonomy, Phylogeny, Molecular Dating and Ancestral State Reconstruction of Xylariomycetidae (Sordariomycetes)
Marker :RTM415,D2000 DNA ladder
Author: Kevin D Hyde,Jian-Kui Liu
Journal: Fungal Diversity volume 112, pages 1–88 (2022)
Institution:School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Paper link:https://link.springer.com/article/10.1007/s13225-021-00495-5
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