{"product_id":"neb-m0315s","title":"New England Biolabs, M0315S, Terminal Transferase","description":"Terminal transferase catalyzes the addition of deoxynucleotides to the 3' hydroxyl terminus of DNA molecules.  \u003cb\u003eRelated Categories\u003c\/b\u003e DNA Labeling,, DNA Manipulation  \u003cb\u003eApplications\u003c\/b\u003e A-tailing,, PCR  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol dTTP into acid-insoluble material in a total reaction volume of 50ul in 1 hour at 37°C using d(A) 18 as primer.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X Terminal Transferase Reaction Buffer Pack Supplement with 0.25 mM CoCl2 Incubate at 37°C 1X Terminal Transferase Reaction Buffer Pack 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate (pH 7.9 @ 25°C)  \u003cb\u003eStorage Buffer\u003c\/b\u003e 50 mM KPO4 100 mM NaCl 1.43 mM β-ME 50% Glycerol 0.1% Triton® X-100 pH 7.3 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 75°C for 20 minutes  \u003cb\u003eMolecular Weight\u003c\/b\u003e Theoretical: 58000 daltons  \u003cb\u003e5' - 3' Exonuclease\u003c\/b\u003e No  \u003cb\u003e3' - 5' Exonuclease\u003c\/b\u003e No  \u003cb\u003eStrand Displacement\u003c\/b\u003e No  \u003cb\u003eUnit Assay Conditions\u003c\/b\u003e 1X Terminal Transferase Reaction Buffer, 0.72 μM d(A) 18 , 0.2 mM dTTP, and 1 μCi [ 3 H]- dTTP in a 50 μl total reaction volume.  \u003cb\u003eFAQ\u003c\/b\u003e Q: Can Terminal Transferase be heat inactivated? A: Yes, heat to 75°C for 20 minutes. Alternatively, reactions can be stopped by adding 10 μl of 0.2 M EDTA. Q: Can Terminal Transferase label the 5' end of DNA? A: No. Use T4 Polynucleotide Kinase (NEB# M0201) to label the 5' end. Q: Is Terminal Transferase supplied with dNTPs? A: No, the dNTPs must be ordered separately. They can be ordered as a set of 4 individual tubes, Deoxynucleotide Solution Set (NEB# N0446) with a 100mM concentration of each nucleotide. Q: Does Terminal Transferase require Co2+ as a divalent cation? A: Co2+ increases the incorporation of pyrimidines (Maniatis), and makes addition to blunt and 3' recessed ends more efficient (Current Protocols). Q: Can just one dNTP be added to the end of DNA with Terminal Transferase? A: Yes, single nucleotides can be added by incorporation of a chain terminating analog such as a dideoxynucleotide. (ddATP) Q: Does Terminal Transferase have a preference for one type of 3' DNA end? A: Addition of dNTPs to 3' OH protruding ends is more efficient than with 3' OH recessed or blunt ends. Q: What are the kM values for Terminal Transferase? A: * 100 μM for dATP * 100 μM for dTTP * 500 μM for dCTP * 500 μM for dGTP * 1 μM for oligos * 1 mM for homopolymer primer ends Q: Will Terminal Transferase add a non-radioactively labeled dNTP? A: Yes, Terminal Transferase will incorporate biotinylated nucleotides. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835480690857,"sku":"M0315S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0315s","provider":"Iright","version":"1.0","type":"link"}