CometAssay Electrophoresis System II

DNA Damage Assay HT1080

Experiment
DNA Damage Assay HT1080
Product
CometAssay Electrophoresis System II from Bio-Techne
Manufacturer
Bio-Techne

Protocol tips

Upstream tips
Cell suspension at a concentration of 10^5cells/ml was mixed in a 1:1 ratio withTrevigen LMAgarose (#4250-050-02) at 37◦C.
Downstream tips
Use Cell Profiler software (version 2.1.1 rev 6c2d896) for analyzing the images of comet.

Publication protocol

After treatment, cells were trypsinized with 0.25% trypsin for several minutes at 37◦C. The trypsin was inactivated with a 4-fold volume of DMEM. Cell suspension at a concentration of 105cells/ml was mixed in a 1:1 ratio withTrevigen LMAgarose (#4250-050-02) at 37◦C. The mixturewas pipetted onto comet slides (Trevigen, #3950-300-02)that had been pre-coated with a 1% normal melting pointagarose (Sigma) base layer. The drop containing the cellswas covered with a glass cover slip and incubated at 4◦Cfor5 min. After incubation, the cover slips were removed, andthe slides were immersed in lysis solution (30 mM ethylene-diaminetetraacetic acid (EDTA), 0.5% sodium dodecyl sul-phate (SDS) and 10 mM Tris–HCl, pH 8.0, supplementedwith 500g/ml proteinase K) and incubated at 37◦Cfor1 h. After lysis, the slides were washed three times for 5min in PBS and incubated in 1×TBE (Tris-Borate-EDTAbuffer) for 20 min at 4◦C. Electrophoresis was performedin a Trevigen electrophoresis system (#4250-050-ES) for 10min at 4◦Cand1V/cm in 1×TBE. The comets were counterstained with SYBR Green for 1 h (1:3000; Thermo Sci-entific, #S7563). The comets were visualized at four magni-fication using an inverted Nikon Eclipse Ti-E fluorescencemicroscope equipped with a Nikon Intensilight C-HGFIlight source (objective: Nikon Plan Fluor 4/0.13; camera:DS-Qi2). The images of the comets were analyzed usingCellProfiler software (version 2.1.1 rev 6c2d896)

Full paper   Login or join for free to view the full paper.

Reviews

CometAssay Electrophoresis System II from Bio-Techne has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA Damage Assay HT1080 using CometAssay Electrophoresis System II from Bio-Techne.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Bio-Techne for CometAssay Electrophoresis System II below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA Damage Assay HT1080 using CometAssay Electrophoresis System II from Bio-Techne. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms