DNA Repair Enzymes and Structure-specific Endonucleases are enzymes which cleave DNA at a specific DNA lesion or structure. (To learn about non-specific endonucleases and exonucleases, visit here.) These enzymes can be used in a wide variety of applications such as:
- Repair of DNA sample degradation due to oxidative damage, UV radiation, ionizing radiation, phenol/chloroform extraction, mechanical shearing, formalin fixation (post extraction) or long term storage
- Base excision repair (BER)
- DNA mismatch repair
- Nucleotide excision repair
- Forensic analysis of environmental samples, analysis of ancient DNA, DNA damage control, and DNA-DNA and protein-DNA interactions
- Preparation for downstream applications such as PCR, microarray analysis, or other DNA technologies
- Single cell gel electrophoresis (Comet assay) to assess samples for DNA damage
- Genotoxicity tests by alkaline elution or alkaline unwinding
- Elimination or repair of large DNA secondary structures using T7 Endonuclease I (NEB #M0302)
- What reaction conditions were used to define Authenticase™?
- How does Authenticase™ improve the quality and fidelity of PCR gene assembly?
- How do I convert my gene of interest into oligonucleotides?
- Why do you recommend setting up two tubes for the PCR reaction containing different amounts of Authenticase™-treated samples as templates?
- Can I use Authenticase™ for genome editing applications?
- Does Authenticase™ recognize single base pair mismatches or indels (insertions/deletions)?
- What common additives inhibit Authenticase™?
- What PCR reagents are recommended for DNA amplification in genome editing (CRISPR/Cas9, TALEN, ZFN) mismatch detection assays?
- Can I use Authenticase™ genome editing (CRISPR/Cas9, TALEN, ZFN) mismatch detection assays with unpurified PCR products?
- What size PCR amplicon should I design to analyze the genomic editing efficiency?
- If my PCR reaction yield is low, can I add more than 5 µl of the PCR reaction to the digestion reaction?
- Why do I see an extra band when I run the undigested heteroduplex on a Bioanalyzer or agarose gel?
- What are the differences between Mismatch Endonuclease I (NEB #M0678) and Authenticase (NEB #M0689)?
- Comet Assay - Modified for Detection of Oxidized Bases Using the Repair Endonucleases Fpg, hOGG1 and Endonuclease III (Nth)
- Control Reaction Protocol for PreCR Repair Mix
- Sequential Reaction Protocol for PreCR Repair Mix
- Standard E. coli DNA Gyrase Reaction (M0306)
- Standard Reaction Protocol for PreCR Repair Mix
- Using RecA and an oligonucleotide to form a stable triple helix
- Determining Genome Targeting Efficiency using T7 Endonuclease I
- T7 Endonuclease I-based Mutation Detection with the EnGen® Mutation Detection Kit (NEB #E3321)
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- Error Correction During Gene Synthesis (NEB #M0689)
- Supplemental Protocol 1: Generation of DNA fragments by PCR assembly of pooled oligos (NEB #M0689)
- Supplemental Protocol 2: Using colony PCR to identify positive clones (NEB #M0689)
- Mismatch Detection Assay (NEB #M0689)
- Transfection of EnGen® Spy Cas9 HF1 (NEB #M0667) into adherent cells using the Lipofectamine® RNAiMAX System
DNA Damage - the major cause of missing pieces from the DNA puzzle
- Activities of DNA Repair Enzymes and Structure-specific Endonucleases
- Properties of DNA Repair Enzymes and Structure-specific Endonucleases
- DNA Damage and PreCR
- Mauris, J.and Evans, T.C., Jr. (2010) A human PMS2 homologue from Aquifex aeolicus stimulates an ATP-dependent DNA helicase. J Biol Chem; 285(15), 11087-11092. PubMedID: 20129926
- Karbaschi, Mahsa., Macip, Salvador., Mistry, Vilas., Abbas, Hussein H.K., Delinassios, George J., Evans, Mark D., Young, Antony R., Cooke, Marcus S. (2015) Rescue of cells from apoptosis increases DNA repair in UVB exposed cells: implications for the DNA damage response Toxicol Res; 4, 725-738. DOI: doiL 10.1039/c4tx00197d
- Gehring, A.M., Zatopek, K.M., Burkhart, B.W., Potapov, V., Santangelo, T.J., Gardner, A.F (2019) Biochemical reconstitution and genetic characterization of the major oxidative damage base excision DNA repair pathway in Thermococcus kodakarensis DNA Repair (Amst); PubMedID: 31841800, DOI: 10.1016/j.dnarep.2019.102767
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Learn about endonucleases and some of their common applications.
This webinar reviews the activities of exonucleases and endonucleases, provides insight on how to choose the right exo- or endonuclease for various molecular biology applications, and explains how to use these reagents when developing new molecular biology workflows.