Homologous Recombination Assay Kit Rapid and sensitive tool for measuring the efficiency of homologous recombination For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Homologous Recombination Assay Kit Rapid and sensitive tool for measuring the efficiency of homologous recombination USD$529.00 SKU 35600 Size 200 Transformations / 24 Transfections Quantity Overview Details Documentation Citations Overview Rapid and sensitive tool for measuring the efficiency of homologous recombinationFor any cell types including prokaryotes and eukaryotesAll primers included for quantification of the homologous recombination efficiency through qPCR. Norgen’s Homologous Recombination Assay Kit provides a rapid and sensitive tool for measuring the efficiency of homologous recombination in both bacterial and mammalian cells. The kit consists of 5 components: (1) dl-1 plasmid, (2) dl-2 plasmid, (3) positive control plasmid, (4) assay primers mixture, and (5) universal primers mixture. The assay is based on the co-transformation of the two dl plasmids into bacterial cells followed by the isolation of plasmid DNA and quantification of the homologous recombination efficiency through qPCR. In addition, PCR can be used for qualitative analysis of the recombination efficiency. The primer mixture provided with the kit is designed to amplify only the homologous recombination product but not the dl plasmids (dl-1 or dl-2). The two plasmids can be co-transfected into mammalian cells, then the DNA can be isolated and the homologous recombination efficiency can be assayed by qPCR (or PCR). In addition, the efficiency of homologous recombination can be assessed in bacteria by blue/white screening using X-gal. The provided positive control contains the functional LacZ sequence and can be used as a positive control for the plasmid DNA isolation efficiency (after transformation or transfection) or PCRs. The use of PCR-based homologous recombination assay enhances the sensitivity and eliminates the need for labor intensive screening by using traditional protein expression and staining techniques and is suitable for high throughput screening studies. The kit is designed to perform 200 transformations or 24 transfections. Details Supporting Data Previous Figure 1. Schematic Representation of Homologous Recombination Assay. The lacZα coding region of the positive control plasmid was mutated and two plasmids with different mutations were generated. Therefore, the two plasmids containing different defective lacZα cassettes can form a functional lacZα cassette through intermolecular homologous recombination. The two defective plasmids, once transformed into bacterial cells, will form white colonies. Only the plasmid that has undergone homologous recombination and generated the functional lacZα can form blue colonies. The homologous recombination efficiencies can be determined by calculating the percentages of the blue colonies among all the blue and white colonies. Figure 2: Real-time PCR Amplification of Plasmid DNA Isolated from HEK 293 Cells. Aftter transfection with the supplied plasmids genomic DNA was isolated from each well of a 6-well plate using Norgen’s RNA/DNA/Protein Purification Kit. Fifty nanograms of the positive control plasmid was transfected into each well of the HEK 293 cells, and the total DNA was isolated 12 h later. For the recombinant samples, 25 ng of dl-1 plus 25 ng of dl-2 were transfected and then isolated. For the negative controls, 50 ng of dl-1 or 50 ng of dl-2 were transfected and then isolated. The experiment was carried out in duplicate to avoid the reading error of the PCR machine. Next Click for expanded view Homologous Recombination Assay Kit Contents: 1. dl-1 Plasmid 2. dl-2 Plasmid 3. Positive Control Plasmid 4. Assay Primers Mixture 5. Universal Primers Mixture 6. Nuclease-Free Water 7. Product Insert SHELF LIFE AND HANDLING All solutions should be kept tightly sealed and stored at -20oC or lower temperature for up to 1 year without any reduction in kit performance. Kit Component Cat. 35600 (200 transformations/24 transfections) dl-1 Plasmid 12 μg (50 ng/μL) dl-2 Plasmid 12 μg (50 ng/μL) Positive Control Plasmid 5 μg (50 ng/μL) Assays Primers Mixture 400 μL Universal Primers Mixture 400 μL Nuclease-Free Water 1.25 mL Product Insert 1 Documentation PROTOCOLS (35600) Homologous Recombination Assay Kit - Protocol (200 transformations/24 transfections) SAFETY DATA SHEETS (35600) Homologous Recombination Assay Kit - Safety Data Sheet Citations Title Blocking lipid synthesis induces DNA damage in prostate cancer and increases cell death caused by PARP inhibition Journal Science Signaling. 2024. Authors Caroline Fidalgo Ribeiro, Silvia Rodrigues, Debora Campanella Bastos, Giuseppe Nicolò Fanelli, Hubert Pakula, Marco Foiani, Giorgia Zadra, Massimo Loda Title Replicative Stress Coincides with Impaired Nuclear DNA Damage Response in COX4-1 Deficiency Journal Int. J. Mol. Sci.. 2022. Authors Liza Douiev , Chaya Miller, Guy Keller, Hadar Benyamini, Bassam Abu-Libdeh and Ann Saada Title Phase I Clinical Trial of DNA Methyltransferase Inhibitor Decitabine and PARP Inhibitor Talazoparib Combination Therapy in Relapsed/Refractory Acute Myeloid Leukemia. Journal Clin Cancer Res. 2022. Authors Maria R. Baer, Aksinija A. Kogan, Søren M. Bentzen, Tian Mi, Rena G. Lapidus, Vu H. Duong, Ashkan Emadi, Sandrine Niyongere, Casey L. O'Connell, Benjamin A. Youngblood, Stephen B. Baylin, Feyruz V. Rassool Title Modulation of BRCA1 mediated DNA damage repair by deregulated ER-α signaling in breast cancers Journal Am J Cancer Res. 2022. Authors Arathi Rajan, Geetu R Varghese, Induprabha Yadev, Jaimie Anandan, Neetha R Latha, Dipyaman Patra, Neethu Krishnan, Krithiga Kuppusamy, Arathy V Warrier, Satej Bhushan, Revathy Nadhan, Ram Mohan Ram Kumar, and Priya Srinivas Title Hepatocyte Growth Factor Improves the Therapeutic Efficacy of Human Bone Marrow Mesenchymal Stem Cells via RAD51 Journal Molecular Therapy. 2018. Authors Eun Ju Lee, Injoo Hwang, Ji Yeon Lee, Jong Nam Park, Keun Cheon Kim, Gi-Hwam Kim, Chang-Mo Kang, Irene Kim, Seo-Yeon Lee, Hyo-Soo Kim Title Significance of cyclin D1 overexpression in progression and radio-resistance of pediatric ependymomas Journal Oncotarget. 2017. Authors Muh-Lii Liang, Tsung-Han Hsieh, Yun-Ru Liu, Yi-Wei Chen, Yi-Yen Lee, Feng-Chi Chang, Shih-Chiej Lin, Ming-Chao Huang, Donald Ming-Tak Ho, Tai-Tong Wong, Yun Yen, and Muh-Hwa Yang Title Trifluoperazine, a novel autophagy inhibitor, increases radiosensitivity in glioblastoma by impairing homologous recombination Journal Journal of Experimental & Clinical Research. 2017. Authors Xin Zhang, Ran Xu, Chao Zhang, Yangyang Xu, Mingzhi Han, Bin Huang, Anjing Chen, Chen Qui, Frits Thorsenm Lars Prestegarden, Rolf Bjerkvig, Jian Wang and Xingang Li Title 14-3-3sigma Contributes to Radioresistance by Regulating DNA Repair and Cell Cycle via PARP1 and CHK2 Journal Molecular Cancer Research. 2017. Authors Chen Y, Li Z, Dong Z, Beebe J, Yang K, Fu L, Zhang JT. Title Effect of endonuclease G depletion on plasmid DNA uptake and levels of homologous recombination in hela cells Journal Molecular Cell Biology. 2016. Authors V. Misic, M. El-Mogy, S. Geng, Y. Haj-Ahmad Title Protein–Protein Interaction Inhibitors of BRCA1 Discovered Using Small Molecule Microarrays Journal Small Molecule Microarrays. 2016. Authors Na Z, Pan S, Uttamchandani M, Yao SQ. Title Hypoxia Differentially Modulates the Genomic Stability of Clinical-Grade ADSCs and BM-MSCs in Long-Term Culture Journal Stem Cells. 2015 Authors Bigot N, Mouche A, Preti M, Loisel S, Renoud ML, Le Guével R, Sensebé L, Tarte K, Pedeux R Title Targeting MPS1 Enhances Radiosensitization of Human Glioblastoma by Modulating DNA Repair Proteins. Journal Molecular Cancer Research. 2015 Authors Maachani UB, Kramp T, Hanson R, Zhao S, Celiku O, Shankavaram U, Colombo R, Caplen NJ, Camphausen K, Tandle A Title Mutant IDH1-driven cellular transformation increases RAD51-mediated homologous recombination and temozolomide resistance. Journal Cancer Research. 2014. Authors Ohba S, Mukherjee J, See WL, Pieper RO. Related Products Plasmid MiniPrep Kit 2X PCR Master Mix
Homologous Recombination Assay Kit Rapid and sensitive tool for measuring the efficiency of homologous recombination For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Homologous Recombination Assay Kit Rapid and sensitive tool for measuring the efficiency of homologous recombination USD$529.00 SKU 35600 Size 200 Transformations / 24 Transfections Quantity