DNA Fiber Labeling and Imaging

Cells were labeled with 50 μM CldU (Sigma), exposed to HU (4 mM) and labeled with 50 μM IdU (Sigma) as indicated in the figures. DNA fibers were spread as previously described (35 (link)), and fiber tracts were detected using anti-IdU (BD Biosciences, 347580) and anti-CldU (Novus Biologicals, NB500-169) primary antibodies and Alexa Fluor 488 and 555 secondary antibodies (Invitrogen). Fibers were imaged at 60× magnification with oil immersion using a Zeiss microscope and analyzed with ImageJ.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Eckelmann B.J., Bacolla A., Wang H., Ye Z., Guerrero E.N., Jiang W., El-Zein R., Hegde M.L., Tomkinson A.E., Tainer J.A, & Mitra S. (2020). XRCC1 promotes replication restart, nascent fork degradation and mutagenic DNA repair in BRCA2-deficient cells. NAR Cancer, 2(3), zcaa013.

Publication 2020

anti-IdU NB500-169 Alexa Fluor 488 and 555

Alexa fluor 488 Antibodies Biologicals Cells Fiber Immersion Microscope Novus

Corresponding Organization :

Other organizations : Houston Methodist, Texas A&M Health Science Center, The University of Texas MD Anderson Cancer Center, Instituto Conmemorativo Gorgas de Estudios de la Salud, Chinese Academy of Medical Sciences & Peking Union Medical College, Cornell University, New Mexico Cancer Center, University of New Mexico

Top 5 similar protocols

Variable analysis

independent variables

Exposure to HU (4 mM)

dependent variables

DNA fiber tracts

control variables

Labeling with 50 μM CldU (Sigma)
Labeling with 50 μM IdU (Sigma)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!