Visualizing DNA replication by single-molecule analysis of replicated DNA.
Visualizing DNA replication by single-molecule analysis of replicated DNA.
复制标题
通过复制的DNA的单分子分析来可视化DNA复制。
DOI：
10.1016/j.xpro.2023.102721
复制
发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Gerhardt, Jeannine
中科院分区：
文献类型：
--
作者：
Madireddy, Advaitha;Gerhardt, Jeannine
复制DNA的单分子分析（SMARD）是一种独特的技术，能够以单分子分辨率可视化特定基因组区域的DNA复制。在这里，我们提出了一个协议，可视化的DNA复制的SMARD。我们描述了脉冲标记DNA的步骤，然后分离和拉伸基因组DNA。然后，我们详细的检测在染色体区域的复制，通过免疫染色和荧光原位杂交。使用SMARD，我们可以可视化复制启动，进展，终止和分叉停滞。有关本方案使用和执行的完整详细信息，请参见Norio等人。（2001）和Gerhardt et al.（2014年）。SMARD揭示了特定的基因组位点是如何复制的这种独特的技术结合了DNA纤维分析与FISH SMARD能够检测复制叉方向和复制叉停滞使用SMARD，可以同时检测起始和终止位点出版商注：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。复制DNA的单分子分析（SMARD）是一种独特的技术，能够以单分子分辨率可视化特定基因组区域的DNA复制。在这里，我们提出了一个协议，可视化的DNA复制的SMARD。我们描述了脉冲标记DNA的步骤，然后分离和拉伸基因组DNA。然后，我们详细的检测在染色体区域的复制，通过免疫染色和荧光原位杂交。使用SMARD，我们可以可视化复制启动，进展，终止和分叉停滞。
Single-molecule analysis of replicated DNA (SMARD) is a unique technique that enables visualization of DNA replication at specific genomic regions at single-molecule resolution. Here, we present a protocol for visualizing DNA replication by SMARD. We describe steps for pulse labeling DNA, followed by isolating and stretching of genomic DNA. We then detail the detection of the replication at chromosomal regions through immunostaining and fluorescence in situ hybridization. Using SMARD, we can visualize replication initiation, progression, termination, and fork stalling. For complete details on the use and execution of this protocol, please refer to Norio et al. (2001) and Gerhardt et al. (2014). SMARD reveals how specific genomic loci are replicated This unique technique combines DNA fiber analysis with FISH SMARD is able to detect replication fork direction and replication fork stalling Using SMARD, initiation and termination sites are detected simultaneously Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Single-molecule analysis of replicated DNA (SMARD) is a unique technique that enables visualization of DNA replication at specific genomic regions at single-molecule resolution. Here, we present a protocol for visualizing DNA replication by SMARD. We describe steps for pulse labeling DNA, followed by isolating and stretching of genomic DNA. We then detail the detection of the replication at chromosomal regions through immunostaining and fluorescence in situ hybridization. Using SMARD, we can visualize replication initiation, progression, termination, and fork stalling.
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影响因子：
9.8
作者：
Demczuk A;Gauthier MG;Veras I;Kosiyatrakul S;Schildkraut CL;Busslinger M;Bechhoefer J;Norio P
通讯作者：
Norio P
影响因子：
16
作者：
Gerhardt, Jeannine;Tomishima, Mark J.;Schildkraut, Carl L.
通讯作者：
Schildkraut, Carl L.
影响因子：
14.9
作者：
Madireddy A;Purushothaman P;Loosbroock CP;Robertson ES;Schildkraut CL;Verma SC
通讯作者：
Verma SC
影响因子：
7.8
作者：
Gerhardt, Jeannine;Zaninovic, Nikica;Schildkraut, Carl L.
通讯作者：
Schildkraut, Carl L.
影响因子：
82.9
作者：
Jasra, Sakshi;Giricz, Orsi;Zeig-Owens, Rachel;Pradhan, Kith;Goldfarb, David G.;Barreto-Galvez, Angelica;Silver, Alexander J.;Chen, Jiahao;Sahu, Srabani;Gordon-Mitchell, Shanisha;Choudhary, Gaurav S.;Aluri, Srinivas;Bhagat, Tushar D.;Shastri, Aditi;Bejan, Cosmin A.;Stockton, Shannon S.;Spaulding, Travis P.;Thiruthuvanathan, Victor;Goto, Hiroki;Gerhardt, Jeannine;Haider, Syed Hissam;Veerappan, Arul;Bartenstein, Matthias;Nwankwo, George;Landgren, Ola;Weiden, Michael D.;Lekostaj, Jacqueline;Bender, Ryan;Fletcher, Frederick;Greenberger, Lee;Ebert, Benjamin L.;Steidl, Ulrich;Will, Britta;Nolan, Anna;Madireddy, Advaitha;Savona, Michael R.;Prezant, David J.;Verma, Amit
通讯作者：
Verma, Amit