{"product_id":"neb-m0304l","title":"New England Biolabs, M0304L, Endonuclease IV","description":"\u003cb\u003eRelated Categories\u003c\/b\u003e DNA Repair Enzymes and Structure-specific Endonucleases  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34-mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 μl in 1 hour at 37°C. *An AP site is created by treating 10 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X NEBuffer™ 3 Incubate at 37°C 1X NEBuffer™ 3 100 mM NaCl 50 mM Tris-HCl 10 mM MgCl2 1 mM DTT (pH 7.9 @ 25°C)  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 250 mM NaCl 1 mM DTT 0.1 mM EDTA 200 µg\/ml BSA 50% Glycerol 0.15% Triton® X-100 pH 7.4 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 85°C for 20 minutes  \u003cb\u003eUnit Assay Conditions\u003c\/b\u003e 1X NEBuffer 3 containing 10 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 μl.  \u003cb\u003eFAQ\u003c\/b\u003e Q: What is the activity of Endonuclease IV in the NEBuffers? A: The Endo IV optimum buffer is 1X NEBuffer 3. It has no activity in NEBuffer 1, 100% in NEBuffer 2 and 25% activity in NEBuffer 4. Q: What is the molecular weight for Endonuclease IV? A: The molecular weight of Endonuclease IV is 31kDa. Q: Does Endonuclease IV cleave ssDNA? A: Yes, Endonuclease IV cleaves abasic sites on both ssDNA and dsDNA. Q: Will the 5' terminus left by an Endonuclease IV cleavage be suitable for subsequent ligation by T4 DNA ligase? A: The 5' end generated by Endonuclease IV cleavage is not suitable for ligation. Q: In addition to AP sites, does Endonuclease IV have cleavage activity on other types of DNA damage? A: Endo IV also recognizes 5,6-dihydrothymine. However, its activity on this substrate is less than on AP sites. For more information please refer to this page in our website: (DNA Repair Enzymes). Q: Can an abasic site be created and then cleaved with Endonuclease IV in the same reaction? How active is Uracil-DNA-Glycosylase (cat# M0280S) in Endonuclease IV buffer? A: The Endonuclease IV buffer is NEBuffer 3. UDG is 100% active in NEBuffer 3 and therefore it is possible to perform both reactions in NEBuffer3. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835528269993,"sku":"M0304L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0304l","provider":"Iright","version":"1.0","type":"link"}