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首頁 > 核酸生物學(xué) > 核酸電泳 > PAGE電泳Marker

10bp DNA ladder(10-100 bp)

10bp DNA ladder(10-100 bp)

產(chǎn)品編號:RTM443

產(chǎn)品規(guī)格:50次

數(shù)量
價(jià)格 ¥500

10bp DNA ladder

● 產(chǎn)品編號及規(guī)格:

RTM443          50次 (250 μl)    

● 產(chǎn)品組成:

貨號

名稱

規(guī)格

RTM443-01

10bp DNA ladder

50次(250 μl

DL070-01

6×Native-PAGE DNA上樣緩沖液

1 ml




● 產(chǎn)品簡介:

本產(chǎn)品是由8條帶狀雙鏈DNA條帶組成的精準(zhǔn)定量Marker,適用于聚丙烯酰胺凝膠電泳中DNA條帶大小和含量的分析。8條帶的大小分別為10,15,20,25,30,40,50,100bp。其中25 bp條帶為加亮帶,含量為100 ng/5 μl,其余條帶濃度為50 ng/5 μl。

本產(chǎn)品為雙鏈DNA Marker,適用于非變性的PAGE電泳和瓊脂糖凝膠電泳,不適用于尿素PAGE電泳。由于片段很小,不建議用瓊脂糖凝膠電泳分離。

按照每次上樣5 μl計(jì)算,該產(chǎn)品可以使用50次。

儲存、效期及運(yùn)輸條件:

  -20℃ 貯存;有效期3年;濕冰運(yùn)輸。

● 使用方法:

一、 TBE-PAGE凝膠分離:

1、制備凝膠步驟:

1.1參照凝膠模具說明書,裝配好凝膠模具。

1.2按照表一將不同體積的成分在小燒杯或試管中混合;最后加入10%APS和TEMED,輕輕攪拌使其混勻,避免產(chǎn)生氣泡。

   注:10 bp DNA ladder 適用于配制20%TBE-PAGE膠。

1.3在凝膠模具中灌入適量分離膠溶液(對于mini-gel,凝膠液加至約距前玻璃板頂端1.5 cm或距梳齒約0.5 cm即可),然后在分離膠溶液上輕輕覆蓋一層1-2 cm的無水乙醇,使凝膠表面保持平整。

1.4靜置10-20分鐘,待分離膠和乙醇層之間出現(xiàn)一個(gè)清晰的界面后,說明凝膠已聚合

表一 TBE-PAGE分離膠配方表 (總體積5 ml,適用于1mm厚度小板膠)

各組份體積(ml

最佳DNA分離范圍

凝膠濃度

滅菌水

30%PAA291

5×TBE

10%APS

TEMED

70-450 bp

6%

3

1.0

1.0

0.05

0.005

60-460 bp

8%

2.66

1.34

1.0

0.05

0.005

50-300 bp

10%

2.33

1.67

1.0

0.05

0.005

40-200 bp

12%

2

2.0

1.0

0.05

0.005

25-150 bp

15%

0.5

2.5

1.0

0.05

0.005

5-100 bp

20%

0.66

3.34

1.0

0.05

0.005

1.5去除覆蓋在分離膠上的乙醇;按照表二將不同體積成分在一個(gè)小燒杯或試管中混合;最后加入10%過硫酸銨和TEMED,輕輕攪拌使其混勻,避免產(chǎn)生氣泡。

表二 TBE-PAGE 4%濃縮配方表 (總體積1.5 ml,適用于1mm厚度小板膠)

各組份體積(ml

凝膠濃度

滅菌水

30%PAA291

5×TBE

10%APS

TEMED

4%

1.0

0.2

0.3

0.02

0.002

1.6將濃縮膠溶液加至分離膠的上面,直至凝膠溶液到達(dá)前玻璃板的頂端;將梳子插入凝膠內(nèi),避免產(chǎn)生氣泡。

1.7靜置30-60分鐘,等待濃縮膠聚合。

注:凝膠的聚合時(shí)間與環(huán)境溫度有關(guān)。夏天溫度較高時(shí),聚合較快;冬天氣溫低時(shí),聚合時(shí)間會延長??梢愿鶕?jù)環(huán)境溫度的不同調(diào)節(jié)APS的加入量。

2、電泳:

2.1 將凝膠板固定在電泳裝置上,往上槽和下槽中加入1×TBE電泳液,1 ml吸頭沖洗加樣孔1-2次。

2.2 取待測樣品,加入相應(yīng)體積6×Native PAGE DNA上樣緩沖液,如5 μl樣品加1 μl 上樣緩沖液,短暫離心后取5-10 μl上樣。 10 bp DNA ladder  1 mm厚10齒梳子上樣5 μl,其余梳齒和厚度凝膠適量調(diào)整上樣量。

2.3 連接電源線,打開電源開關(guān)。200 V穩(wěn)壓電泳,至溴酚藍(lán)指示前沿距離玻璃下沿1 cm時(shí)結(jié)束電泳。

TBE-PAGE電泳

恒電壓

起始電流

結(jié)束電流

電泳時(shí)間

200 V

20-25 mA/板膠

10-15 mA/板膠

50+min


          20%T3.3C TBE-PAGE

3、染色:

3.1 漂洗:玻璃板上拆下凝膠后,適量蒸餾水漂洗3-5分鐘。

3.2 染色液配制:

  TBE-PAGE膠可以使用EBRealSafe類染料后染染色,以下程序用RealSafe Red核酸染料(貨號:GR002)進(jìn)行染色。即用型染色液配制:

 

即用型RealSafe核酸染色液

 

100 ml

1×TBE

100 ml

RealSafe Red核酸染料

20 μl

3.3染色和觀察:

  凝膠浸泡于即用型染色液中,常溫避光搖床40-60 rpm染色30 分鐘。紫外光下觀察。

4、實(shí)驗(yàn)示例:


20TBE-PAGE電泳

左1-7:10,15,20,25,30,40,50 bp

右1:10 bp DNA ladder

凝膠長度8 cm

電壓200 V 起始電流25 mA,終止電流13 mA

電泳緩沖液:1×TBE

電泳時(shí)間:50分鐘

染色:RealSafe Red核酸染料后染30分鐘


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 linkhttps://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 linkhttps://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 oneJanuary 2012 | Volume 7 | Issue 1 | e29875

InstitutionState 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

InstitutionSchool of Agriculture and Biology, Shanghai Jiao Tong University

Paper linkhttps://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

InstitutionKey Laboratory of Crop Genetics and Breeding of Hainan Province, Hainan Academy of Agricultural Sciences

Paper linkhttps://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 OnePublished: December 21, 2015

InstitutionKey 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

InstitutionState Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences

Paper linkhttps://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

InstitutionState Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences

Paper linkhttps://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

InstitutionKey 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

InstitutionInstitute 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

InstitutionState 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

InstitutionBeijing Research Center for Agricultural Standards and Testing, Beijing Academy of Agriculture and Forestry Sciences

Paper linkhttps://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)

InstitutionSchool of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China

Paper linkhttps://link.springer.com/article/10.1007/s13225-021-00495-5

 

14. [2021 IF=8.025] A method for high-concentration agarose gel preparation and its

application in high-resolution separation of low-molecular-weight nucleic acids and proteins.

Marker :RTM443,10bp DNA ladder

Author: Lili Chang , Dan Wang, Cunzhi Peng, Qi Wang, Bingqiang Xu, Zheng Tong

Journal: International Journal of Biological Macromolecules 231 (2023) 123358

InstitutionKey Laboratory of Biology and Genetic Resources of Tropical Crops, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural

Sciences

Paper linkhttps://doi.org/10.1016/j.ijbiomac.2023.123358

 


 
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中文字幕人妻少妇伦伦AV99久久人妻无码精品系列_最新高清无码专区无套内谢少妇毛片免费看看我出血_一色一伦一区二区三区色欲AV亚洲中文字幕久久_精品一区二区三区在线成人漫画无套内谢少妇毛片免费看看我出血