pRYG is a pUC19 derivative in which the 206 bp SphI-NarI fragment of pUC19 was replaced with a 245 bp SphI-AccI fragment upstream of the human adult beta-globin gene promoter. The 245 bp fragment contains a 54 bp alternating repeat of purine/pyrimidine sequence that constitutes a strong eukaryotic topo II cleavage site. Cleavages within the repeat are readily detected in the absence of topo II inhibitors such as m-AMSA or VM-26. The sequence of the plasmid is given in Table 2 of Spitzner et al.Description of Product
- Topotecan Hydrochloride (Hycamtin)
- Camptothecin
- 9-Nitro-20 (S) Camptothecin (Rubitecan)
- 10-Hydroxy Camptothecin
- 9-Amino Camptothecin
- Doxorubicin (Adriacin)
- Daunomycin
- Etoposide (VP16)
- Amsacrine (m-AMSA)
- Genistein
- Ellipticine
- HU-331
Gyrase & Topo Iv
- Ciprofloxacin
- Topoisomerase II Alpha C-Terminal Peptide
- Polyclonal Antibody to Human Topoisomerase II Alpha
- Monoclonal Antibody to Human Topoisomerase II Alpha
- Polyclonal Antibody to Human Topoiosomerase II Beta
- Polyclonal Antibody to Yeast Topoisomerase II (Serum)
- Monoclonal Antibody to Human Topoiosomerase II Beta
- Topoisomerase II Immunoflourescence Kit
- Topoisomerase II Immunoprecipitation Kit
- Topoisomerase II Band Shift Kit
Cell Context DNA Repair
- Human: HeLa Reporter Cell Line (Includes I-Sce1 Transfection)
- Human: HeLa HR Reporter Cell line (Includes CRISPR-CAS Transfection)
- Human: Neuroblastoma HR Reporter Cell line (SH-SY5Y, Includes I-Sce1 Transfection)
- Mouse: 3T3 HR Reporter Cell Line (Includes I-Sce1 Transfection)
- Mouse: Neuronal Lineage HR Reporter Cell line (Includes I-Sce1 Transfection)
- Custom Cell Based Screening Kit for (HR) Pathway
- Human:HeLa NHEJ Reporter Cell Line (Includes I-Sce1 Transfection)
- Human:HNSH NHEJ Reporter Cell line (Includes I-Sce1 Transfection)
- Mouse:3T3 Fibroblast NHEJ Reporter (Includes I-Sce1 Transfection)
- Custom Cell Based Screening Kit for NHEJ
Topo Enzymes
DNA Gyrases
Topo Kits
Drug Screening Kits
Topo Antibodies
Protein Mw Marker & Antiobody Kits
DNA Substrates
TDP1 Products
TDP2 Products
Topo Related Drugs
Stable Cell Lines for DNA Repair Assays
Transfection-based DNA Repair Assays
Topo Enzymes
DNA Gyrases
Topo Kits
Drug Screening Kits
Topo Antibodies
Protein Mw Marker & Antiobody Kits
DNA Substrates
Top2/Gyrase Substrates & Markers
Top1 pHOT-1 DNA Substrates & Markers
TDP1 Products
TDP2 Products
Topo Related Drugs
Stable Cell Lines for DNA Repair Assays
These HR/NHEJ kits are cell based, meaning they have the HR or NHEJ GFP reporters pre-installed. A single plasmid (I-Sce1) is transfected to activate repair. Please see this table for a summary and comparison/application of each kit.
Homologous GFP Repair Systems
HeLa Cell HR Kit (I-Sce1 Based)
HeLa Cell HR Kit (CRISPR-CAS Based)
Human Neuroblastoma Cell Line SH-SY5Y using (I-Sce1 transfection)
Mouse 3T3 Cell Line using I-Sce1 transfection
Mouse Neuronal Cell Line using I-Sce1 transfection
Custom construction Kit to make your own HR Cell Reporter
NHEJ GFP Reporter
Systems
Transfection-based DNA Repair Assays
Transfection-based HR and NHEJ kits use transient transfections to make a pool of cells containing the DNA reporters to allow the investigator to interrogate the 2 major DNA repair pathways. The advantages are:
- It uses GFP readout to measure the efficiency of repair.
- There is zero background when repair is blocked or inhibited
- The reporters for HR or NHEJ can be installed in virtually any cell line that can be transiently transfect
- It is rapid and robust
- It is ideal for drug screens to find novel inhibitors of HR/NHEJ
- The kits are highly specific and can only report HR or NHEJ
- Readout GFP can be done by FACS or Imaging
- Entire pathway can be followed by live imaging in viable cells
- Ideal to track repair kinetics at the single cell level
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