{"product_id":"neb-m0269l","title":"New England Biolabs, M0269L, phi29 DNA Polymerase","description":"Perfect for multiple displacement amplification  \u003cb\u003eRelated Categories\u003c\/b\u003e Isothermal Amplification \u0026amp; Strand Displacement  \u003cb\u003eApplications\u003c\/b\u003e Whole Genome Amplification \u0026amp; Multiple Displacement Amplification,, Isothermal Amplification  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme that will incorporate 0.5 pmol of dNTP into acid insoluble material in 10 minutes at 30°C.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X phi29 DNA Polymerase Reaction Buffer Supplement with Recombinant Albumin, Molecular Biology Grade Incubate at 30°C 1X phi29 DNA Polymerase Reaction Buffer 50 mM Tris-HCl 10 mM MgCl2 10 mM (NH4)2SO4 4 mM DTT (pH 7.5 @ 25°C)  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 100 mM KCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 0.5% Tween® 20 0.5% IGEPAL® CA-630 pH 7.4 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 65°C for 10 minutes  \u003cb\u003eMolecular Weight\u003c\/b\u003e Theoretical: 67000 daltons  \u003cb\u003e5' - 3' Exonuclease\u003c\/b\u003e No  \u003cb\u003e3' - 5' Exonuclease\u003c\/b\u003e Yes  \u003cb\u003eStrand Displacement\u003c\/b\u003e ++++  \u003cb\u003eUnit Assay Conditions\u003c\/b\u003e 1X phi29 DNA Polymerase Reaction Buffer, 0.1 mg\/ml Recombinant Albumin, 0.01 mg\/ml HindIII-digested λ DNA, 0.2 µM dTTP including [ 3 H]-dTTP, 0.2 mM dGTP, 0.2 mM dATP and 0.2 mM dCTP.  \u003cb\u003eFAQ\u003c\/b\u003e Q: Can phi29 DNA Polymerase be used in other NEBuffers? A: Yes. Phi29 DNA Polymerase is 30% active in NEBuffer 1, 35% active in NEBuffer 2, 25% active in NEBuffer 3, 90% active in NEBuffer 4, and 30% active in ThermoPol Buffer. It also is 100% active in rCutSmart buffer with the addition of 4mM DTT (final concentration). Q: Can phi29 DNA Polymerase be used to blunt DNA? A: It produces blunt-end fragments during DNA synthesis, so in theory it could be used to blunt DNA. Traditionally, DNA Polymerase I, Large (Klenow) Fragment (NEB# M0210) or T4 DNA Polymerase (NEB# M0203) are used to blunt DNA. Q: Can phi29 DNA Polymerase be used to fill in 3' overhangs? A: No, DNA polymerases cannot fill in 3' overhangs. To create blunt ends 3' overhangs must be removed. DNA Polymerase I, Large (Klenow) Fragment (NEB# M0210) or T4 DNA Polymerase (NEB# M0203) are the best choices to remove 3' overhangs. Mung Bean Nuclease (NEB# M0250) can also be used to remove 3´ overhangs. Q: Can phi29 DNA Polymerase be used to remove 5' overhangs? A: No, DNA polymerases cannot remove 5' overhangs. Use Mung Bean Nuclease (NEB# M0250) to remove 5' overhangs. Q: Can phi29 DNA Polymerase be heat inactivated? A: Yes, heat at 65°C for 10 minutes. Q: At what rate does phi29 DNA Polymerase add nucleotides to a primed single-stranded template? A: The following are phi29 DNA Polymerase rates at different temperatures using a primed single-stranded M13 template(1): 2280 nt\/min at 30°C 1490 nt\/min at 15°C 290 nt\/mn at 4°C (1) Soengas, M.S., Gutierrez, M.S. and Salas, M. 1995 J. Mol. Biol.253(4):517-529. Q: Will phi29 DNA Polymerase work in rCutSmart buffer? A: Yes, phi29 DNA Polymerase will work in rCutSmart buffer. To see its % functional activity in rCutSmart, and that of other DNA Modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart. Q: Are NEB DNA Polymerases supplied with dNTPs? A: No, the dNTPs must be ordered separately. They can be ordered as a convenient mix (Deoxynucleotide (dNTP) Solution Mix (NEB# N0447) with a 10 mM concentration of each deoxynucleotide) or as a set of 4 individual tubes (Deoxynucleotide (dNTP) Solution Set (NEB# N0446) with a 100mM concentration of each deoxynucleotide). ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835540328617,"sku":"M0269L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0269l","provider":"Iright","version":"1.0","type":"link"}