您好,歡迎光(guāng)臨中科(kē)瑞泰!

免費咨詢電話(huà):
400-699-0631

首頁 > 核酸生(shēng)物學 > 核酸電泳 > PAGE電泳Marker

10bp DNA ladder(10-100 bp)

10bp DNA ladder(10-100 bp)

産品編号:RTM443

産品規格:50次

數量
價格 ¥500

10bp DNA ladder

● 産品編号及規格:

RTM443          50次 (250 μl)    

● 産品組成:

貨号

名稱

規格

RTM443-01

10bp DNA ladder

50次(250 μl

DL070-01

6×Native-PAGE DNA上樣緩沖液

1 ml




● 産品簡介:

本産品是由8條帶狀雙鏈DNA條帶組成的精準定量Marker,适用于聚丙烯酰胺凝膠電泳中DNA條帶大(dà)小和含量的分(fēn)析。8條帶的大(dà)小分(fēn)别爲10,15,20,25,30,40,50,100bp。其中25 bp條帶爲加亮帶,含量爲100 ng/5 μl,其餘條帶濃度爲50 ng/5 μl。

本産品爲雙鏈DNA Marker,适用于非變性的PAGE電泳和瓊脂糖凝膠電泳,不适用于尿素PAGE電泳。由于片段很小,不建議(yì)用瓊脂糖凝膠電泳分(fēn)離(lí)。

按照(zhào)每次上樣5 μl計(jì)算,該産品可(kě)以使用50次。

儲存、效期及運輸條件(jiàn):

  4℃ 貯存6個月(-20℃有效期3年(nián));濕冰運輸。

● 使用方法:

一、 TBE-PAGE凝膠分(fēn)離(lí):

1、制備凝膠步驟:

1.1參照(zhào)凝膠模具說(shuō)明書(shū),裝配好凝膠模具。

1.2按照(zhào)表一将不同體(tǐ)積的成分(fēn)在小燒杯或試管中混合;最後加入10%APS和TEMED,輕輕攪拌使其混勻,避免産生(shēng)氣泡。

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

1.3在凝膠模具中灌入适量分(fēn)離(lí)膠溶液(對于mini-gel,凝膠液加至約距前玻璃闆頂端1.5 cm或距梳齒約0.5 cm即可(kě)),然後在分(fēn)離(lí)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(jìng)置10-20分(fēn)鍾,待分(fēn)離(lí)膠和乙醇層之間出現一個清晰的界面後,說(shuō)明凝膠已聚合

表一 TBE-PAGE分(fēn)離(lí)膠配方表 (總體(tǐ)積5 ml,适用于1mm厚度小闆膠)

各組份體(tǐ)積(ml

最佳DNA分(fēn)離(lí)範圍

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(qù)除覆蓋在分(fēn)離(lí)膠上的乙醇;按照(zhào)表二将不同體(tǐ)積成分(fēn)在一個小燒杯或試管中混合;最後加入10%過硫酸铵和TEMED,輕輕攪拌使其混勻,避免産生(shēng)氣泡。

表二 TBE-PAGE 4%濃縮配方表 (總體(tǐ)積1.5 ml,适用于1mm厚度小闆膠)

各組份體(tǐ)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将濃縮膠溶液加至分(fēn)離(lí)膠的上面,直至凝膠溶液到達前玻璃闆的頂端;将梳子插入凝膠内,避免産生(shēng)氣泡。

1.7靜(jìng)置30-60分(fēn)鍾,等待濃縮膠聚合。

注:凝膠的聚合時間與環境溫度有關。夏天溫度較高時,聚合較快(kuài);冬天氣溫低時,聚合時間會延長。可(kě)以根據環境溫度的不同調節APS的加入量。

2、電泳:

2.1 将凝膠闆固定在電泳裝置上,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(xiàng)應體(tǐ)積6×Native PAGE DNA上樣緩沖液,如(rú)5 μl樣品加1 μl 上樣緩沖液,短(duǎn)暫離(lí)心後取5-10 μl上樣。 10 bp DNA ladder  1 mm厚10齒梳子上樣5 μl,其餘梳齒和厚度凝膠适量調整上樣量。

2.3 連接電源線,打開電源開關。200 V穩壓電泳,至溴酚藍指示前沿距離(lí)玻璃下沿1 cm時結束電泳

TBE-PAGE電泳

恒電壓

起始電流

結束電流

電泳時間

200 V

20-25 mA/闆膠

10-15 mA/闆膠

50+min


          20%T3.3C TBE-PAGE

3、染色:

3.1 漂洗:玻璃闆上拆下凝膠後,适量蒸餾水漂洗3-5分(fēn)鍾。

3.2 染色液配制:

  TBE-PAGE膠可(kě)以使用EBRealSafe類染料後染染色,以下程序用RealSafe Red核酸染料(貨号:GR002)進行染色。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(guāng)搖床40-60 rpm染色30 分(fēn)鍾。紫外光(guāng)下觀察。

4、實驗示例:


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

電泳時間:50分(fēn)鍾

染色:RealSafe Red核酸染料後染30分(fēn)鍾


使用DNA Marker産品發表部分(fēn)文章(zhāng)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 oneSeptember 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 Biology14 (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: Phytotaxa2015,227 (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

 


 
隻有購(gòu)買過該商品的用戶才能進行評價。[發表評價]