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pbr hpv18  (New England Biolabs)


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    Structured Review

    New England Biolabs pbr hpv18
    A. qPCR was performed to measure <t>HPV18</t> DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).
    Pbr Hpv18, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 3184 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pbr+hpv18/NcoI/pmc07263449-693-16-36
    Average 98 stars, based on 3184 article reviews
    pbr hpv18 - by Bioz Stars, 2026-09
    98/100 stars

    Images

    1) Product Images from "Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes"

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    Journal: Current protocols in microbiology

    doi: 10.1002/cpmc.101

    A. qPCR was performed to measure HPV18 DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).
    Figure Legend Snippet: A. qPCR was performed to measure HPV18 DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).

    Techniques Used:

    Quasivirus stocks were digested with nuclease to remove DNA outside of the capsid and the viral DNA was extracted and quantified by qPCR for HPV18 DNA. Shown are examples of viral genome equivalents (VGE) per μl obtained from quasivirus stocks prepared with Basic Protocol 1: Transfection, Harvest, and Isolation of HPV Quasiviruses; Alternate Protocol 1: Packaging HPV DNA Replicated in 293TT Cells; Support Protocol 1: Production of HPV Minicircles and Alternate Support Protocol 1: Production of Recircularized HPV Genomes.
    Figure Legend Snippet: Quasivirus stocks were digested with nuclease to remove DNA outside of the capsid and the viral DNA was extracted and quantified by qPCR for HPV18 DNA. Shown are examples of viral genome equivalents (VGE) per μl obtained from quasivirus stocks prepared with Basic Protocol 1: Transfection, Harvest, and Isolation of HPV Quasiviruses; Alternate Protocol 1: Packaging HPV DNA Replicated in 293TT Cells; Support Protocol 1: Production of HPV Minicircles and Alternate Support Protocol 1: Production of Recircularized HPV Genomes.

    Techniques Used: Transfection, Isolation

    A. Map of pMC.BESPX-HPV18
    Figure Legend Snippet: A. Map of pMC.BESPX-HPV18

    Techniques Used:

    A. Map of pUC-HPV18 showing the NcoI sites used to remove the vector and recircularize the viral genome.
    Figure Legend Snippet: A. Map of pUC-HPV18 showing the NcoI sites used to remove the vector and recircularize the viral genome.

    Techniques Used: Plasmid Preparation

    A. Duplicate wells of HFKs were infected with HPV18 Quasivirus (prepared by Basic Protocol 1) at an MOI of 100. RNA was collected at 24, 48, 72, and 96 hpi. RT-qPCR for viral transcripts E1Ê4 (left) and E6*I (right) was performed to quantitate viral transcription. Error bars represent the standard deviation of replicate wells.
    Figure Legend Snippet: A. Duplicate wells of HFKs were infected with HPV18 Quasivirus (prepared by Basic Protocol 1) at an MOI of 100. RNA was collected at 24, 48, 72, and 96 hpi. RT-qPCR for viral transcripts E1Ê4 (left) and E6*I (right) was performed to quantitate viral transcription. Error bars represent the standard deviation of replicate wells.

    Techniques Used: Infection, Quantitative RT-PCR, Standard Deviation

    Related Articles

    Plasmid Preparation:

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. Materials list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Polymerase Chain Reaction:

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. Materials list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Purification:

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. Materials list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Blocking Assay:

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. Materials list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Agarose Gel Electrophoresis:

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. Materials list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes
    Article Snippet: .. list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI. ..



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    pbr hpv18  (New England Biolabs)
    98
    New England Biolabs pbr hpv18
    A. qPCR was performed to measure <t>HPV18</t> DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).
    Pbr Hpv18, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pbr+hpv18/NcoI/pmc07263449-693-16-36
    Average 98 stars, based on 1 article reviews
    pbr hpv18 - by Bioz Stars, 2026-09
    98/100 stars
    		  Buy from Supplier

    Image Search Results


    A. qPCR was performed to measure HPV18 DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).

    Journal: Current protocols in microbiology

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    doi: 10.1002/cpmc.101

    Figure Lengend Snippet: A. qPCR was performed to measure HPV18 DNA in fractions prepared by standard (Basic Protocol 1) and Ripcord (Alternative Protocol 2).

    Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI.

    Techniques:

    Quasivirus stocks were digested with nuclease to remove DNA outside of the capsid and the viral DNA was extracted and quantified by qPCR for HPV18 DNA. Shown are examples of viral genome equivalents (VGE) per μl obtained from quasivirus stocks prepared with Basic Protocol 1: Transfection, Harvest, and Isolation of HPV Quasiviruses; Alternate Protocol 1: Packaging HPV DNA Replicated in 293TT Cells; Support Protocol 1: Production of HPV Minicircles and Alternate Support Protocol 1: Production of Recircularized HPV Genomes.

    Journal: Current protocols in microbiology

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    doi: 10.1002/cpmc.101

    Figure Lengend Snippet: Quasivirus stocks were digested with nuclease to remove DNA outside of the capsid and the viral DNA was extracted and quantified by qPCR for HPV18 DNA. Shown are examples of viral genome equivalents (VGE) per μl obtained from quasivirus stocks prepared with Basic Protocol 1: Transfection, Harvest, and Isolation of HPV Quasiviruses; Alternate Protocol 1: Packaging HPV DNA Replicated in 293TT Cells; Support Protocol 1: Production of HPV Minicircles and Alternate Support Protocol 1: Production of Recircularized HPV Genomes.

    Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI.

    Techniques: Transfection, Isolation

    A. Map of pMC.BESPX-HPV18

    Journal: Current protocols in microbiology

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    doi: 10.1002/cpmc.101

    Figure Lengend Snippet: A. Map of pMC.BESPX-HPV18

    Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI.

    Techniques:

    A. Map of pUC-HPV18 showing the NcoI sites used to remove the vector and recircularize the viral genome.

    Journal: Current protocols in microbiology

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    doi: 10.1002/cpmc.101

    Figure Lengend Snippet: A. Map of pUC-HPV18 showing the NcoI sites used to remove the vector and recircularize the viral genome.

    Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI.

    Techniques: Plasmid Preparation

    A. Duplicate wells of HFKs were infected with HPV18 Quasivirus (prepared by Basic Protocol 1) at an MOI of 100. RNA was collected at 24, 48, 72, and 96 hpi. RT-qPCR for viral transcripts E1Ê4 (left) and E6*I (right) was performed to quantitate viral transcription. Error bars represent the standard deviation of replicate wells.

    Journal: Current protocols in microbiology

    Article Title: Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes

    doi: 10.1002/cpmc.101

    Figure Lengend Snippet: A. Duplicate wells of HFKs were infected with HPV18 Quasivirus (prepared by Basic Protocol 1) at an MOI of 100. RNA was collected at 24, 48, 72, and 96 hpi. RT-qPCR for viral transcripts E1Ê4 (left) and E6*I (right) was performed to quantitate viral transcription. Error bars represent the standard deviation of replicate wells.

    Article Snippet: list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 HPV genome such as pUC-HPV18 ( Cole & Danos, 1987 ) or pBR-HPV18 ( Boshart et al., 1984 ) (HPV genomes can be acquired from The HPV Reference Center) NcoI (50,000 U/ml) (NEB #R0193T), EcoRI (20,000 U/ml) (NEB# R0101L) or other enzyme to release viral genome from the bacterial vector NEB CutSmart Buffer (NEB #B7204S) QIAquick PCR Purification Kit (Qiagen #28104) Gel Loading Dye, Purple (6X) (NEB #B7024S) SeaKem GTG Agarose (Lonza #50074) Ethidium Bromide (Invitrogen #15585011) TAE Buffer (Tris-acetate-EDTA) (50X) (Thermo Fisher #B49) T4 DNA ligase (400,000 U/ml) (NEB #M0202S) 10X T4 DNA Ligase Reaction Buffer (NEB #B0202S) Isopropanol (Sigma #190764) 5M NaCl (KD Medical #RGF-3270) 70% Ethanol TE pH 8.0 (Quality Biological #351-011-131) Heat block capable of cooling such as ThermoMixer C (Eppendorf #5382000023) or 16°C water bath in a cold room Refrigerated tabletop centrifuge such as Centrifuge 5415R (Eppendorf #5401000137) Agarose gel running apparatus list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Prepare a 50 μl restriction digestion reaction to free HPV18 from the bacterial plasmid backbone containing 10 μg pUC-HPV18, 1X CutSmart Buffer, and 50 U NcoI.

    Techniques: Infection, Quantitative RT-PCR, Standard Deviation