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首页 > 核酸生物学 > 核酸电泳 > PAGE电泳Marker

10bp DNA ladder(10-100 bp)

10bp DNA ladder(10-100 bp)

产品编号:RTM443

产品规格:50次

数量
价格 ¥500

10bp DNA ladder

● 产品编号及规格:

RTM443          50次 (250 μl)    

● 产品组成:

货号

名称

规格

RTM443-01

10bp DNA ladder

50次(250 μl

DL070-01

6×Native-PAGE DNA上样缓冲液

1 ml




● 产品简介:

本产品是由8条带状双链DNA条带组成的精准定量Marker,适用于聚丙烯酰胺凝胶电泳中DNA条带大小和含量的分析。8条带的大小分别为10,15,20,25,30,40,50,100bp。其中25 bp条带为加亮带,含量为100 ng/5 μl,其余条带浓度为50 ng/5 μl。

本产品为双链DNA Marker,适用于非变性的PAGE电泳和琼脂糖凝胶电泳,不适用于尿素PAGE电泳。由于片段很小,不建议用琼脂糖凝胶电泳分离。

按照每次上样5 μl计算,该产品可以使用50次。

储存、效期及运输条件:

  4℃ 贮存6个月(-20℃有效期3年);湿冰运输。

● 使用方法:

一、 TBE-PAGE凝胶分离:

1、制备凝胶步骤:

1.1参照凝胶模具说明书,装配好凝胶模具。

1.2按照表一将不同体积的成分在小烧杯或试管中混合;最后加入10%APS和TEMED,轻轻搅拌使其混匀,避免产生气泡。

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

1.3在凝胶模具中灌入适量分离胶溶液(对于mini-gel,凝胶液加至约距前玻璃板顶端1.5 cm或距梳齿约0.5 cm即可),然后在分离胶溶液上轻轻覆盖一层1-2 cm的无水乙醇,使凝胶表面保持平整。

1.4静置10-20分钟,待分离胶和乙醇层之间出现一个清晰的界面后,说明凝胶已聚合

表一 TBE-PAGE分离胶配方表 (总体积5 ml,适用于1mm厚度小板胶)

各组份体积(ml

最佳DNA分离范围

凝胶浓度

灭菌水

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去除覆盖在分离胶上的乙醇;按照表二将不同体积成分在一个小烧杯或试管中混合;最后加入10%过硫酸铵和TEMED,轻轻搅拌使其混匀,避免产生气泡。

表二 TBE-PAGE 4%浓缩配方表 (总体积1.5 ml,适用于1mm厚度小板胶)

各组份体积(ml

凝胶浓度

灭菌水

30%PAA291

5×TBE

10%APS

TEMED

4%

1.0

0.2

0.3

0.02

0.002

1.6将浓缩胶溶液加至分离胶的上面,直至凝胶溶液到达前玻璃板的顶端;将梳子插入凝胶内,避免产生气泡。

1.7静置30-60分钟,等待浓缩胶聚合。

注:凝胶的聚合时间与环境温度有关。夏天温度较高时,聚合较快;冬天气温低时,聚合时间会延长。可以根据环境温度的不同调节APS的加入量。

2、电泳:

2.1 将凝胶板固定在电泳装置上,往上槽和下槽中加入1×TBE电泳液,1 ml吸头冲洗加样孔1-2次。

2.2 取待测样品,加入相应体积6×Native PAGE DNA上样缓冲液,如5 μl样品加1 μl 上样缓冲液,短暂离心后取5-10 μl上样。 10 bp DNA ladder  1 mm厚10齿梳子上样5 μl,其余梳齿和厚度凝胶适量调整上样量。

2.3 连接电源线,打开电源开关。200 V稳压电泳,至溴酚蓝指示前沿距离玻璃下沿1 cm时结束电泳

TBE-PAGE电泳

恒电压

起始电流

结束电流

电泳时间

200 V

20-25 mA/板胶

10-15 mA/板胶

50+min


          20%T3.3C TBE-PAGE

3、染色:

3.1 漂洗:玻璃板上拆下凝胶后,适量蒸馏水漂洗3-5分钟。

3.2 染色液配制:

  TBE-PAGE胶可以使用EBRealSafe类染料后染染色,以下程序用RealSafe Red核酸染料(货号:GR002)进行染色。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染色和观察:

  凝胶浸泡于即用型染色液中,常温避光摇床40-60 rpm染色30 分钟。紫外光下观察。

4、实验示例:


20TBE-PAGE电泳

左1-7:10,15,20,25,30,40,50 bp

右1:10 bp DNA ladder

凝胶长度8 cm

电压200 V 起始电流25 mA,终止电流13 mA

电泳缓冲液:1×TBE

电泳时间:50分钟

染色:RealSafe Red核酸染料后染30分钟


使用DNA Marker产品发表部分文章列表:

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 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

 


 
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