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Single Molecular Resolution to Monitor DNA Replication Fork Dynamics upon Stress by DNA Fiber Assay

Author: Wenpeng Liu, date: 12/20/2021, view: 1217, Q&A: 0

[Abstract]

DNA replication always encounters numerous endogenous and exogenous stresses during the cell cycle. Measuring the cell responses to stress has primarily relied on cell survival and incorporation of radioactive dNTPs, which is limited in resolution. A higher resolution is required to monitor how replication and repair respond to these stresses.

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Assembly of Genetic Circuits with the Mammalian ToolKit

Authors: João P. Fonseca, Alain R Bonny, Jason Town and Hana El-Samad, date: 03/05/2020, view: 3896, Q&A: 0

[Abstract] The ability to rapidly assemble and prototype cellular circuits is vital for biological research and its applications in biotechnology and medicine. The Mammalian ToolKit (MTK) is a Golden Gate-based cloning toolkit for fast, reproducible and versatile assembly of large DNA vectors and their implementation in mammalian models. The MTK consists of ...

Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120

Authors: Amit Srivastava, Anand Ballal, Karl Forchhammer and Anil Kumar Tripathi, date: 02/20/2020, view: 3704, Q&A: 0

[Abstract] Anabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a ...

Evaluation of Genotoxicity by Micronucleus Assay in vitro and by Allium cepa Test in vivo

Authors: Christina N. Banti and Sotiris K. Hadjikakou, date: 07/20/2019, view: 6343, Q&A: 0

[Abstract] The in vitro and in vivo genotoxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) is evaluated by the micronucleus test and the Allium cepa assay, respectively. Fetal lung fibroblast cells (MRC-5), normal human corneal epithelial cells (HCEC) and ...

DNA Fiber Assay upon Treatment with Ultraviolet Radiations

Authors: Alfano Luigi, Antonio Giordano and Francesca Pentimalli, date: 06/05/2017, view: 17097, Q&A: 1

[Abstract] Genome stability is continuously challenged by a wide range of DNA damaging factors. To promote a correct DNA repair and cell survival, cells orchestrate a coordinated and finely tuned cascade of events collectively known as the DNA Damage Response (DDR). Ultra Violet (UV) rays are among the main environmental sources of DNA damage and a well ...

Single-molecule Analysis of DNA Replication Dynamics in Budding Yeast and Human Cells by DNA Combing

Authors: Hélène Tourrière, Julie Saksouk, Armelle Lengronne and Philippe Pasero , date: 06/05/2017, view: 12012, Q&A: 0

[Abstract] The DNA combing method allows the analysis of DNA replication at the level of individual DNA molecules stretched along silane-coated glass coverslips. Before DNA extraction, ongoing DNA synthesis is labeled with halogenated analogues of thymidine. Replication tracks are visualized by immunofluorescence using specific antibodies. Unlike biochemical ...

Cloud-point PEG Glass Surfaces for Imaging of Immobilized Single Molecules by Total-internal-reflection Microscopy

Authors: Zhengjian Zhang, Sang Ryul Park, Alexandros Pertsinidis and Andrey Revyakin, date: 04/05/2016, view: 10204, Q&A: 0

[Abstract] This effective, robust protocol generates glass coverslips coated with biotin-functionalized polyethylene glycol (PEG), making the glass surface resistant to non-specific absorption of biomolecules, and permitting immobilization of biomolecules for subsequent single-molecule tracking of biochemical reactions. The protocol can be completed in one ...

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