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packagene lambda dna packaging system  (Promega)

 
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    Promega packagene lambda dna packaging system
    Visualizing unwinding of individual <t>DNA</t> molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated <t>lambda</t> DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.
    Packagene Lambda Dna Packaging System, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packagene lambda dna packaging system/product/Promega
    Average 90 stars, based on 1 article reviews
    packagene lambda dna packaging system - by Bioz Stars, 2026-02
    90/100 stars

    Images

    1) Product Images from "Direct Fluorescent Imaging of Translocation and Unwinding by Individual DNA Helicases"

    Article Title: Direct Fluorescent Imaging of Translocation and Unwinding by Individual DNA Helicases

    Journal: Methods in enzymology

    doi: 10.1016/bs.mie.2016.09.010

    Visualizing unwinding of individual DNA molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated lambda DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.
    Figure Legend Snippet: Visualizing unwinding of individual DNA molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated lambda DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.

    Techniques Used: Microscopy, Staining, Construct, Lambda DNA Preparation, Injection, Binding Assay



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    Visualizing unwinding of individual <t>DNA</t> molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated <t>lambda</t> DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.
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    Visualizing unwinding of individual <t>DNA</t> molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated <t>lambda</t> DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.
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    Visualizing unwinding of individual <t>DNA</t> molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated <t>lambda</t> DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.
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    Image Search Results


    Visualizing unwinding of individual DNA molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated lambda DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.

    Journal: Methods in enzymology

    Article Title: Direct Fluorescent Imaging of Translocation and Unwinding by Individual DNA Helicases

    doi: 10.1016/bs.mie.2016.09.010

    Figure Lengend Snippet: Visualizing unwinding of individual DNA molecules using fluorescent SSB protein and TIRF microscopy. (A) Top: Diagram of phage λ DNA molecule with the indicated number of biotin groups incorporated in each 12-nt cos overhang. Middle: Illustration of biotinylated λ DNA attached at both ends via biotin–streptavidin linkage. Bottom: Image of an actual λ DNA molecule attached to the glass surface, stained with YO-PRO-1 (100 nM), and illuminated with a 488 nm laser. The image is false colored in green and the attachment points to the glass surface are indicated. (B) The process required to construct a flow cell containing three separate single-channels. The steps highlighted are equivalent to those described in Section 4.1. (C) The flow cell from (B) mounted onto the objective; biotinylated lambda DNA was injected under buffer flow, permitting attachment of both ends to the surface. Unwinding tracks are visualized by binding of AF488-SSBG26C (green) to ssDNA regions. (D) Schematic representation of a TIRF microscope capable of visualizing DNA unwinding by RecQ by monitoring signal from both DNA and fluorescent SSB simultaneously. As shown in (C) the flow cell is mounted onto a 100× oil-immersion objective. The fluorescent SSB and DNA are excited by two lasers; 488 and 561 nm, respectively, and emission measured, via dichroic mirrors (M1 and M2). The deconvoluted emission is then directed onto different areas of a CCD camera generating a signal corresponding to either SSB or DNA. The lasers are operated using a custom LABview VI program to coordinate excitation with image acquisition so that the sample is illuminated only during the exposure times. Panels (A) and (D): From Rad, B., Forget, A. L., Baskin, R. J., & Kowalczykowski, S. C. (2015). Single-molecule visualization of RecQ helicase reveals DNA melting, nucleation, and assembly are required for processive DNA unwinding. Proceedings of the National Academy of Sciences of the United States of America, 112(50), E6852–E6861. http://dx.doi.org/10.1073/pnas.1518028112.

    Article Snippet: The ligated recombinant λ DNA product is packaged into a λ capsid in vitro using the Packagene ® Lambda DNA Packaging System (Promega, Madison, WI).

    Techniques: Microscopy, Staining, Construct, Lambda DNA Preparation, Injection, Binding Assay