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    New England Biolabs nebuffer 3
    HBV capsid assembly in RRL under different conditions The WT HBc proteins were translated in RRL, and the translation reaction mixtures were resolved by agarose gel (1%) electrophoresis (A) or SDS-PAGE (12.5%) (B) without any further treatment (Input) (lane 1) or were treated with <t>NEBuffer</t> 3 alone overnight at 37 °C (Mock) (lane 2), with NEBuffer 3 plus CIAP overnight at 37 °C (CIAP) (lane 3), with NEBuffer 3 plus CIAP overnight at 37 °C followed by RNase treatment for one additional hour (CIAP + RNase) (lane 4), with RNase for 1 h followed by NEBuffer 3 plus CIAP overnight at 37 °C (RNase + CIAP) (lane 5), or with the mixture of phosphatase inhibitors overnight at 37 °C (PPI) (lane 6). All lanes contained 2 µl translation products. 35 )
    Nebuffer 3, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 974 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nebuffer 3/product/New England Biolabs
    Average 99 stars, based on 974 article reviews
    Price from $9.99 to $1999.99
    nebuffer 3 - by Bioz Stars, 2020-08
    99/100 stars
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    HBV capsid assembly in RRL under different conditions The WT HBc proteins were translated in RRL, and the translation reaction mixtures were resolved by agarose gel (1%) electrophoresis (A) or SDS-PAGE (12.5%) (B) without any further treatment (Input) (lane 1) or were treated with NEBuffer 3 alone overnight at 37 °C (Mock) (lane 2), with NEBuffer 3 plus CIAP overnight at 37 °C (CIAP) (lane 3), with NEBuffer 3 plus CIAP overnight at 37 °C followed by RNase treatment for one additional hour (CIAP + RNase) (lane 4), with RNase for 1 h followed by NEBuffer 3 plus CIAP overnight at 37 °C (RNase + CIAP) (lane 5), or with the mixture of phosphatase inhibitors overnight at 37 °C (PPI) (lane 6). All lanes contained 2 µl translation products. 35 )

    Journal: Bio-protocol

    Article Title: Host-regulated Hepatitis B Virus Capsid Assembly in a Mammalian Cell-free System

    doi: 10.21769/BioProtoc.2813

    Figure Lengend Snippet: HBV capsid assembly in RRL under different conditions The WT HBc proteins were translated in RRL, and the translation reaction mixtures were resolved by agarose gel (1%) electrophoresis (A) or SDS-PAGE (12.5%) (B) without any further treatment (Input) (lane 1) or were treated with NEBuffer 3 alone overnight at 37 °C (Mock) (lane 2), with NEBuffer 3 plus CIAP overnight at 37 °C (CIAP) (lane 3), with NEBuffer 3 plus CIAP overnight at 37 °C followed by RNase treatment for one additional hour (CIAP + RNase) (lane 4), with RNase for 1 h followed by NEBuffer 3 plus CIAP overnight at 37 °C (RNase + CIAP) (lane 5), or with the mixture of phosphatase inhibitors overnight at 37 °C (PPI) (lane 6). All lanes contained 2 µl translation products. 35 )

    Article Snippet: Pipette tips (Denville Scientific, catalog numbers: P1096-FR, P1121, P1122, P1126) 1.5 ml microcentrifuge tube (Denville Scientific, catalog number: C2170) Gloves and lab coat (Denville Scientific, catalog number: G4162; Medline Industries, catalog number: 83044QHW) Proteinase K, RNA grade (Thermo Fisher Scientific, Invitrogen™, catalog number: 25530049) Sodium dodecyl sulfate (SDS) (Sigma-Aldrich, catalog number: L4509-1KG) Phenol solution (Sigma-Aldrich, catalog number: P4557) Chloroform (Fisher Scientific, catalog number: BP1145-1) Sodium acetate (Sigma-Aldrich, catalog number: S2889-1KG) Ethyl alcohol (EtOH) (AmericanBio, catalog number: AB00515-00100) UltraPure™ DNase/RNase-Free Distilled Water (Thermo Fisher Scientific, Invitrogen™, catalog number: 10977015) Agarose (Thermo Fisher Scientific, Invitrogen™, catalog number: 15510027) TNT® Coupled Rabbit Reticulocyte Lysate (RRL) (Promega, catalog number: L4610) EasyTag™ L-[35 S]-Methionine (PerkinElmer, catalog number: NEG709A001MC) RNasin® Plus Ribonuclease Inhibitor (Promega, catalog number: N2611) RNaseZap™ RNase Decontamination Solution (Thermo Fisher Scientific, Invitrogen™, catalog number: AM9780) RNase A, DNase and protease-free (Thermo Fisher Scientific, Thermo Scientific™, catalog number: EN0531) NEBuffer 3 (New England Biolabs, catalog number: B7003S) Alkaline Phosphatase, Calf Intestinal (CIAP) (New England Biolabs, catalog number: M0290S) Tris Base (Fisher Scientific, catalog number: BP152-10) Ethylenediaminetetraacetic acid, EDTA (Sigma-Aldrich, catalog number: E5134) Sodium fluoride (Sigma-Aldrich, catalog number: S7920) Sodium pyrophosphate tetrabasic decahydrate (Sigma-Aldrich, catalog number: S6422) β-Glycerophosphate (Sigma-Aldrich, catalog number: G6251) Sodium orthovanadate (Sigma-Aldrich, catalog number: S6508) cOmplete™, EDTA-free Protease Inhibitor Cocktail (Roche Diagnostics, catalog number: 04693132001) TE buffer (see Recipes) 10× phosphatase inhibitors (PPI) (see Recipes) 25× protease inhibitor (see Recipes)

    Techniques: Agarose Gel Electrophoresis, Electrophoresis, SDS Page