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RTM441 20bp DNA ladder(20-500 bp)
 产品货号: RTM441
 产品名称: RTM441 20bp DNA ladder(20-500 bp)
 产品价格/规格: 50次 500元
 产品说明书: 点击查看
 更新时间: 2022-07-29
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  • 20bp DNA ladder

    ● 产品编号及规格:

    RTM441          50次 (250 μl)    

    ● 产品组成:

    货号

    名称

    规格

    RTM441-01

    20bp DNA ladder

    50次(250 μl

    DL070

    6×Native-PAGE DNA上样缓冲液

    1 ml

    DL020

    6×DualColor DNA loading buffer (双染料)

    1 ml

    ● 产品简介:

    本产品是由13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。13条带的大小分别为20,40,60,80,100,120,140,160,180,200,300,400,500 bp。其中100bp和200bp条带为加亮带,含量为100ng/5μl,其余条带浓度为50ng/5μl。

    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。由于片段较小,如使用琼脂糖分离,建议使用高分辨率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。

    ● 储存条件:

      4 贮存6个月(-20℃长期贮存)。

    ● 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:20bp 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。 20bp 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。200V稳压电泳。至二甲苯菁指示前沿到达凝胶三分之二位置时结束电泳(此时溴酚蓝指示前沿已经跑出凝胶,不可见)。

    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

    70+min



    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。

    二、 琼脂糖凝胶分离:

    1. 琼脂糖凝胶制备:

      由于片段较小,建议选用高分辨率琼脂糖(货号AR1036)制胶。

    以下步骤以制备50 ml 3%凝胶为例:

    称取1.5 g琼脂糖于玻璃三角瓶中,加入50 ml 1×TAE,5 μl RealSafe Red核酸染料或其他核酸染料(如EB,GoldView),混匀,盖好瓶盖,微波炉中火加热至沸腾,轻摇混匀,重复1-2次至琼脂糖完全溶解,无可见颗粒。倒入制胶容器中,插好梳子,常温凝固30-50分钟至凝胶完全凝固。

    2. 电泳:

    取待测样品,加入相应体积6×DualColor DNA loading buffer,如10 μl样品加2 μl 上样缓冲液,短暂离心后取5-10 μl上样。 20bp 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。

    建议电泳条件:凝胶浓度为3%,电泳电压8-10 v/cm(单位电压指电泳槽阴阳极之间的距离电压,如阴极阳极之间的距离为20 cm,可以用160-200 V电压进行稳压电泳),待溴酚蓝指示前沿距离凝胶末端2 cm时终止电泳,电泳时间35-40分钟。

    注:3% 琼脂糖TAE电泳中,溴酚蓝大小约为20bp,二甲苯菁大小约200bp。

    3. 紫外灯下观察条带。

    注:使用EB或Goldview染料时,由于染料本身带正电荷较多,随着电泳时间的延长,染料会向阴极聚集,导致小片段核酸结合的染料含量降低,会出现小片段的DNA片段紫外灯下亮度变弱或不可见。此时可以将胶浸泡于含有染料的1×TAE缓冲液中染色15-20分钟即可看到小片段。

    4. 实验示例:



     


  • 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] Identification 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


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