pcr analysis  (Qiagen)


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    QIAamp MinElute Virus Spin Kit
    Description:
    For simultaneous purification of viral DNA and RNA from plasma serum and cell free body fluids Kit contents Qiagen QIAamp MinElute Virus Spin Kit 50 preps 200L Sample 20 to 150L Elution Volume Serum Plasma Sample Viral DNA and RNA Purification Silica Technology Manual Processing MinElute Columns Format For Simultaneous Purification of Viral DNA and RNA from Plasma Serum and Cell free Body Fluids Ideal for PCR Real time PCR Application Includes 50 QIAamp MinElute Columns Qiagen Protease Carrier RNA Buffers 2mL Collection Tubes Benefits Rapid purification of high quality viral DNA and RNA No organic extraction or alcohol precipitation Consistent high yields Complete removal of contaminants and inhibito
    Catalog Number:
    57704
    Price:
    278
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    QIAamp MinElute Virus Spin Kit
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    Structured Review

    Qiagen pcr analysis
    QIAamp MinElute Virus Spin Kit
    For simultaneous purification of viral DNA and RNA from plasma serum and cell free body fluids Kit contents Qiagen QIAamp MinElute Virus Spin Kit 50 preps 200L Sample 20 to 150L Elution Volume Serum Plasma Sample Viral DNA and RNA Purification Silica Technology Manual Processing MinElute Columns Format For Simultaneous Purification of Viral DNA and RNA from Plasma Serum and Cell free Body Fluids Ideal for PCR Real time PCR Application Includes 50 QIAamp MinElute Columns Qiagen Protease Carrier RNA Buffers 2mL Collection Tubes Benefits Rapid purification of high quality viral DNA and RNA No organic extraction or alcohol precipitation Consistent high yields Complete removal of contaminants and inhibito
    https://www.bioz.com/result/pcr analysis/product/Qiagen
    Average 99 stars, based on 6340 article reviews
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    pcr analysis - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle"

    Article Title: Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2018.06.001

    Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.
    Figure Legend Snippet: Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.

    Techniques Used: Selection, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation, Dot Blot, Immunofluorescence, Staining, Purification

    2) Product Images from "Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle"

    Article Title: Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2018.06.001

    Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.
    Figure Legend Snippet: Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.

    Techniques Used: Selection, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation, Dot Blot, Immunofluorescence, Staining, Purification

    3) Product Images from "Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals"

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0067-3

    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)
    Figure Legend Snippet: Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Techniques Used: Real-time Polymerase Chain Reaction, RNA Extraction

    4) Product Images from "Performance of the digene LQ, RH and PS HPVs genotyping systems on clinical samples and comparison with HC2 and PCR-based Linear Array"

    Article Title: Performance of the digene LQ, RH and PS HPVs genotyping systems on clinical samples and comparison with HC2 and PCR-based Linear Array

    Journal: Infectious Agents and Cancer

    doi: 10.1186/1750-9378-6-23

    Type specific concordance between LA and LQ, RH and PS techniques (N = 305) . The genotyping results for each of the tested techniques vs. LA are presented in a color-coded way. Concordances LA/LQ are presented in black, LA/RH in red and LA/PS in blue.
    Figure Legend Snippet: Type specific concordance between LA and LQ, RH and PS techniques (N = 305) . The genotyping results for each of the tested techniques vs. LA are presented in a color-coded way. Concordances LA/LQ are presented in black, LA/RH in red and LA/PS in blue.

    Techniques Used:

    5) Product Images from "Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle"

    Article Title: Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2018.06.001

    Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep . The resultant library production plasmid contained AAV2 ITRs and a modified AAV2 3′ UTR. The AAV library was packaged using standard production protocols as done previously, 41 dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.
    Figure Legend Snippet: Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep . The resultant library production plasmid contained AAV2 ITRs and a modified AAV2 3′ UTR. The AAV library was packaged using standard production protocols as done previously, 41 dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.

    Techniques Used: Selection, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation, Modification, Dot Blot, Immunofluorescence, Staining, Purification

    6) Product Images from "Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals"

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0067-3

    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)
    Figure Legend Snippet: Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Techniques Used: Real-time Polymerase Chain Reaction, RNA Extraction

    7) Product Images from "Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals"

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0067-3

    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)
    Figure Legend Snippet: Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Techniques Used: Real-time Polymerase Chain Reaction, RNA Extraction

    8) Product Images from "Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle"

    Article Title: Bioengineered Viral Platform for Intramuscular Passive Vaccine Delivery to Human Skeletal Muscle

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2018.06.001

    Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep . The resultant library production plasmid contained AAV2 ITRs and a modified AAV2 3′ UTR. The AAV library was packaged using standard production protocols as done previously, 41 dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.
    Figure Legend Snippet: Directed Evolution of AAV Capsids by DNA Shuffling and Selection Screening on Primary Human Skeletal Muscle Stem Cells and Myotubes (A) AAV capsid genes from ten parental serotypes (1, 2, 3b, 4, 5, 6, 8, 9_hu14, avian, and bovine) were PCR-amplified, fragmented with DNaseI digestion, and then randomly reassembled through self-priming PCR. Resultant shuffled Cap genes were cloned back into a replication-competent AAV production plasmid via flanking SwaI/NsiI sites downstream of AAV2 Rep . The resultant library production plasmid contained AAV2 ITRs and a modified AAV2 3′ UTR. The AAV library was packaged using standard production protocols as done previously, 41 dot blot titered, and used for selection. (B) Sequential gating scheme Panels [P]1–P4) to isolate CD31 − CD34 − CD45 − ITGB1 + EGFR + primary human skeletal muscle stem cells from surgical specimens. Numbers indicate percentage of total events falling within each gate. (C) Phase contrast and immunofluorescence PAX7 (pink) and DAPI (blue) nuclear staining of purified primary human muscle stem cell cultures. Scale bar, 100 μm. (D) Diagram illustrating the two selection screens with replicating AAV capsid libraries from (A) that was performed on both pooled primary human skeletal muscle stem cells and human myotubes. (E) Semiquantitative AAV Cap PCR (2.2-Kb product) was performed at each round of each selection screen to demonstrate active replication of AAV library genomes throughout each round of the screen.

    Techniques Used: Selection, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation, Modification, Dot Blot, Immunofluorescence, Staining, Purification

    9) Product Images from "Efficacy and Mechanisms of Murine Norovirus Inhibition by Pulsed-Light Technology"

    Article Title: Efficacy and Mechanisms of Murine Norovirus Inhibition by Pulsed-Light Technology

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.03840-14

    Inactivation of MNV-1 on experimentally contaminated polyethylene (A), polyvinyl chloride (B), and stainless steel (C) disks (1.04 cm 2 ) under clean (bars with light cross-hatching) and fouled (bars with dark cross-hatching) conditions after treatment
    Figure Legend Snippet: Inactivation of MNV-1 on experimentally contaminated polyethylene (A), polyvinyl chloride (B), and stainless steel (C) disks (1.04 cm 2 ) under clean (bars with light cross-hatching) and fouled (bars with dark cross-hatching) conditions after treatment

    Techniques Used:

    SDS-PAGE analysis of purified MNV-1 untreated, treated with 2.07 J cm −2 (3 pulses), or treated with 8.98 J cm −2 (13 pulses). Viral proteins were analyzed via 10% SDS-PAGE followed by Coomassie staining. VP1, major capsid protein.
    Figure Legend Snippet: SDS-PAGE analysis of purified MNV-1 untreated, treated with 2.07 J cm −2 (3 pulses), or treated with 8.98 J cm −2 (13 pulses). Viral proteins were analyzed via 10% SDS-PAGE followed by Coomassie staining. VP1, major capsid protein.

    Techniques Used: SDS Page, Purification, Staining

    Activity of pulsed light against MNV-1 in aqueous suspension.
    Figure Legend Snippet: Activity of pulsed light against MNV-1 in aqueous suspension.

    Techniques Used: Activity Assay

    Viral genomic RNA degraded by pulsed light. Viral genomic RNA extracted from treated or untreated purified MNV-1 was loaded onto a 6000 RNA Pico chip and analyzed on a Bioanalyzer with a UV detector. Results are presented in electropherogram form. Light
    Figure Legend Snippet: Viral genomic RNA degraded by pulsed light. Viral genomic RNA extracted from treated or untreated purified MNV-1 was loaded onto a 6000 RNA Pico chip and analyzed on a Bioanalyzer with a UV detector. Results are presented in electropherogram form. Light

    Techniques Used: Purification, Chromatin Immunoprecipitation

    Viral particles disrupted by pulsed light. Infectivity of purified MNV-1 dropped by 5.15 ± 0.18 log 10 after treatment with 2.07 J cm −2 (3 pulses) and was completely eliminated after treatment with 8.98 J cm −2 (13 pulses). Treated
    Figure Legend Snippet: Viral particles disrupted by pulsed light. Infectivity of purified MNV-1 dropped by 5.15 ± 0.18 log 10 after treatment with 2.07 J cm −2 (3 pulses) and was completely eliminated after treatment with 8.98 J cm −2 (13 pulses). Treated

    Techniques Used: Infection, Purification

    Inactivation of MNV-1 in experimentally contaminated hard waters (A) and turbid waters (B) after pulsed-light treatments at 80 mm from a xenon lamp. White bars, 0.69 J cm − ; white bars with black dots, 2.07 J cm −2 ; checkered bars, 3.45
    Figure Legend Snippet: Inactivation of MNV-1 in experimentally contaminated hard waters (A) and turbid waters (B) after pulsed-light treatments at 80 mm from a xenon lamp. White bars, 0.69 J cm − ; white bars with black dots, 2.07 J cm −2 ; checkered bars, 3.45

    Techniques Used:

    10) Product Images from "An adenovirus-derived protein: A novel candidate for anti-diabetic drug development"

    Article Title: An adenovirus-derived protein: A novel candidate for anti-diabetic drug development

    Journal: Biochimie

    doi: 10.1016/j.biochi.2015.12.002

    E4orf1 transiently but reproducibly enhances glucose excursion: C57BL/6J (9 week old) mice on 60% fat diet since 6 week of age were inoculated with pBabe-puro retrovirus (control; black solid lines, n = 6), or with pBabe expressing E4orf1 (black dotted
    Figure Legend Snippet: E4orf1 transiently but reproducibly enhances glucose excursion: C57BL/6J (9 week old) mice on 60% fat diet since 6 week of age were inoculated with pBabe-puro retrovirus (control; black solid lines, n = 6), or with pBabe expressing E4orf1 (black dotted

    Techniques Used: Mouse Assay, Expressing

    Longer duration of high fat diet delays the improvement in GTT induced by E4orf1: Fourteen week old C57BL/6J male mice on 60% fat diet (since 6wk) were inoculated with pBabe-puro (control; black solid lines/bars, n = 6), or with different doses of pBabe
    Figure Legend Snippet: Longer duration of high fat diet delays the improvement in GTT induced by E4orf1: Fourteen week old C57BL/6J male mice on 60% fat diet (since 6wk) were inoculated with pBabe-puro (control; black solid lines/bars, n = 6), or with different doses of pBabe

    Techniques Used: Mouse Assay

    11) Product Images from "HIV-1 Tat Interacts with a Kaposi’s Sarcoma-Associated Herpesvirus Reactivation-Upregulated Antiangiogenic Long Noncoding RNA, LINC00313, and Antagonizes Its Function"

    Article Title: HIV-1 Tat Interacts with a Kaposi’s Sarcoma-Associated Herpesvirus Reactivation-Upregulated Antiangiogenic Long Noncoding RNA, LINC00313, and Antagonizes Its Function

    Journal: Journal of Virology

    doi: 10.1128/JVI.01280-19

    Soluble HIV Tat enhances the invasion and viral production of KSHV lytic infection-reactivated SLK cells. (A) Viability of control and iSLK-BAC16 cells treated with Dox (1 μg/ml) with or without of HIV Tat (0.2 μg/ml) at the indicated time points was assessed using MTT. (B and D) Representative images from cell migration (B) and invasion (D) assays in iSLK-BAC16 cells treated as described for panel A. (C and E) Quantification of cell migration (C) and invasion (E) described in the legend for panels B and D, respectively. (F) Supernatants from iSLK-BAC16 cells treated as described in the legend for panel A were collected at 72 and 96 h, and the viral titers were determined by analyzing the virion-associated DNA levels using TaqMan qPCR. Data shown are means ± standard deviations (SD) ( n = 3). **, P
    Figure Legend Snippet: Soluble HIV Tat enhances the invasion and viral production of KSHV lytic infection-reactivated SLK cells. (A) Viability of control and iSLK-BAC16 cells treated with Dox (1 μg/ml) with or without of HIV Tat (0.2 μg/ml) at the indicated time points was assessed using MTT. (B and D) Representative images from cell migration (B) and invasion (D) assays in iSLK-BAC16 cells treated as described for panel A. (C and E) Quantification of cell migration (C) and invasion (E) described in the legend for panels B and D, respectively. (F) Supernatants from iSLK-BAC16 cells treated as described in the legend for panel A were collected at 72 and 96 h, and the viral titers were determined by analyzing the virion-associated DNA levels using TaqMan qPCR. Data shown are means ± standard deviations (SD) ( n = 3). **, P

    Techniques Used: Infection, MTT Assay, Migration, Real-time Polymerase Chain Reaction

    12) Product Images from "Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas"

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas

    Journal: EMBO Molecular Medicine

    doi: 10.1002/emmm.201302796

    H-1PV/VPA co-treatment leads to complete remission of established HeLa tumours Source data is available for this figure in the Supporting Information. Analysis of tumour growth. Animals ( n = 8 per group) were treated with either PBS (control), VPA or a combination of both agents as described in Materials and Methods Section. Other animal groups are presented in Supporting Information Fig S7 . The data shown represent average tumour volumes with standard deviation bars. Graphical summary of Kaplan–Meier survival analysis of tumour-bearing nude rats treated as indicated. Animals were sacrificed when the tumour reached the maximum tolerable size of 4000 mm 3 according to law. The difference between H-1PV/VPA co-treatment and treatment with H-1PV alone was statistically significant ( p = 0.0002 as calculated with the two-sided log-rank test and adjusted for multiple testing with the Bonferroni method). Other results from the statistical analysis are shown in Supporting Information Table S2. H-1PV/VPA co-treatment induces DNA damage and apoptosis in vivo . Two rats of each treatment group were sacrificed on day 19. Tumours were fixed, sectioned and examined morphologically either by H E staining or by immunochemistry using antibodies against NS1, VP capsid proteins, γ-H2AX (a DNA damage marker), or active cleaved caspase-3 (an apoptosis marker). Scale bar = 50 µm. Higher levels of NS1, VP1 and VP2 viral proteins in tumour samples from VPA co-treated animals. Tumour-bearing animals ( n = 3) treated with H-1PV alone (total virus dose of 3.6 × 10 9 Vg/animal corresponding to 1.5 × 10 8 pfu/animals subdivided into two subdoses given at days 0 and 7) or co-treated with H-1PV and VPA (100 mg/kg) were sacrificed at day 14. Complete tumours were resected, homogenised and lysed. 20 µg of total cellular extracts were analysed by SDS–PAGE for the presence of the viral proteins. Actin was used as a loading control. VPA-enhanced virus multiplication in vivo . Viral particles were recovered from aliquots of tumour extracts from animals treated as in panel D, purified by using the QIAamp MinElute Virus Spin Kit (Qiagen) and quantified by real time qPCR. Columns represent the average amounts of the virus recovered from the tumours of three animals with standard deviation bars. Numbers on top of the columns indicate the fold increase in virus titers in the presence versus absence of VPA.
    Figure Legend Snippet: H-1PV/VPA co-treatment leads to complete remission of established HeLa tumours Source data is available for this figure in the Supporting Information. Analysis of tumour growth. Animals ( n = 8 per group) were treated with either PBS (control), VPA or a combination of both agents as described in Materials and Methods Section. Other animal groups are presented in Supporting Information Fig S7 . The data shown represent average tumour volumes with standard deviation bars. Graphical summary of Kaplan–Meier survival analysis of tumour-bearing nude rats treated as indicated. Animals were sacrificed when the tumour reached the maximum tolerable size of 4000 mm 3 according to law. The difference between H-1PV/VPA co-treatment and treatment with H-1PV alone was statistically significant ( p = 0.0002 as calculated with the two-sided log-rank test and adjusted for multiple testing with the Bonferroni method). Other results from the statistical analysis are shown in Supporting Information Table S2. H-1PV/VPA co-treatment induces DNA damage and apoptosis in vivo . Two rats of each treatment group were sacrificed on day 19. Tumours were fixed, sectioned and examined morphologically either by H E staining or by immunochemistry using antibodies against NS1, VP capsid proteins, γ-H2AX (a DNA damage marker), or active cleaved caspase-3 (an apoptosis marker). Scale bar = 50 µm. Higher levels of NS1, VP1 and VP2 viral proteins in tumour samples from VPA co-treated animals. Tumour-bearing animals ( n = 3) treated with H-1PV alone (total virus dose of 3.6 × 10 9 Vg/animal corresponding to 1.5 × 10 8 pfu/animals subdivided into two subdoses given at days 0 and 7) or co-treated with H-1PV and VPA (100 mg/kg) were sacrificed at day 14. Complete tumours were resected, homogenised and lysed. 20 µg of total cellular extracts were analysed by SDS–PAGE for the presence of the viral proteins. Actin was used as a loading control. VPA-enhanced virus multiplication in vivo . Viral particles were recovered from aliquots of tumour extracts from animals treated as in panel D, purified by using the QIAamp MinElute Virus Spin Kit (Qiagen) and quantified by real time qPCR. Columns represent the average amounts of the virus recovered from the tumours of three animals with standard deviation bars. Numbers on top of the columns indicate the fold increase in virus titers in the presence versus absence of VPA.

    Techniques Used: Standard Deviation, In Vivo, Staining, Marker, SDS Page, Purification, Real-time Polymerase Chain Reaction

    13) Product Images from "Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals"

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals

    Journal: NPJ Vaccines

    doi: 10.1038/s41541-018-0067-3

    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)
    Figure Legend Snippet: Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Techniques Used: Real-time Polymerase Chain Reaction, RNA Extraction

    14) Product Images from "A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis"

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis

    Journal: Yonsei Medical Journal

    doi: 10.3349/ymj.2012.53.1.132

    Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.

    Techniques Used: Sequencing, Modification, Mutagenesis

    Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.

    Techniques Used: Amplification, Modification, Mutagenesis

    15) Product Images from "A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis"

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis

    Journal: Yonsei Medical Journal

    doi: 10.3349/ymj.2012.53.1.132

    Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.

    Techniques Used: Sequencing, Modification, Mutagenesis

    Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.

    Techniques Used: Amplification, Modification, Mutagenesis

    16) Product Images from "Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection"

    Article Title: Evaluation of the Broad-Range PCR-Electrospray Ionization Mass Spectrometry (PCR/ESI-MS) System and Virus Microarrays for Virus Detection

    Journal: Viruses

    doi: 10.3390/v6051876

    Determination of retrovirus LOD using RT-PCR assays. Total RNA was extracted from each virus dilution to create XMRV and SFV-1 RNA panels (10 0 –10 −7 ) in the absence and presence of 10 5 or 10 4 cell equivalents of Sf9 total nucleic acids as described in Materials and Methods. A subset of the RNA panels (10 −3 –10 −6 ) was subjected to nested RT-PCR assays. An RT minus (−RT) control of the samples shows the absence of residual cellular DNA and the negative, no template control for the PCR shows the absence of contamination in the assay. ( a ) XMRV gag primers; ( b ) SFV-1 gag primers. The size of fragments in the 100 bp marker (M) is indicated in base pairs (bp).
    Figure Legend Snippet: Determination of retrovirus LOD using RT-PCR assays. Total RNA was extracted from each virus dilution to create XMRV and SFV-1 RNA panels (10 0 –10 −7 ) in the absence and presence of 10 5 or 10 4 cell equivalents of Sf9 total nucleic acids as described in Materials and Methods. A subset of the RNA panels (10 −3 –10 −6 ) was subjected to nested RT-PCR assays. An RT minus (−RT) control of the samples shows the absence of residual cellular DNA and the negative, no template control for the PCR shows the absence of contamination in the assay. ( a ) XMRV gag primers; ( b ) SFV-1 gag primers. The size of fragments in the 100 bp marker (M) is indicated in base pairs (bp).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Marker

    17) Product Images from "An adenovirus-derived protein: A novel candidate for anti-diabetic drug development"

    Article Title: An adenovirus-derived protein: A novel candidate for anti-diabetic drug development

    Journal: Biochimie

    doi: 10.1016/j.biochi.2015.12.002

    E4orf1 transiently but reproducibly enhances glucose excursion: C57BL/6J (9 week old) mice on 60% fat diet since 6 week of age were inoculated with pBabe-puro retrovirus (control; black solid lines, n = 6), or with pBabe expressing E4orf1 (black dotted
    Figure Legend Snippet: E4orf1 transiently but reproducibly enhances glucose excursion: C57BL/6J (9 week old) mice on 60% fat diet since 6 week of age were inoculated with pBabe-puro retrovirus (control; black solid lines, n = 6), or with pBabe expressing E4orf1 (black dotted

    Techniques Used: Mouse Assay, Expressing

    A booster dose of E4orf1 does not extend the duration of enhancement in glucose clearance: Nine week old C57BL/6J (9 wk old) male mice on 60% HF diet since 6 wk of age were inoculated with control retrovirus (black solid lines, n = 6), or with E4orf1
    Figure Legend Snippet: A booster dose of E4orf1 does not extend the duration of enhancement in glucose clearance: Nine week old C57BL/6J (9 wk old) male mice on 60% HF diet since 6 wk of age were inoculated with control retrovirus (black solid lines, n = 6), or with E4orf1

    Techniques Used: Mouse Assay

    E4orf1 improves glucose clearance without increasing insulin sensitivity, production or secretion: Insulin tolerance test (ITT) was performed 1 and 2 week post initial inoculation of control retrovirus (black solid lines, n = 3), or with E4orf1 (black
    Figure Legend Snippet: E4orf1 improves glucose clearance without increasing insulin sensitivity, production or secretion: Insulin tolerance test (ITT) was performed 1 and 2 week post initial inoculation of control retrovirus (black solid lines, n = 3), or with E4orf1 (black

    Techniques Used:

    18) Product Images from "A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis"

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis

    Journal: Yonsei Medical Journal

    doi: 10.3349/ymj.2012.53.1.132

    Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Sequencing maps of known EGFR deletion in exon 19 of plasma cfDNA. cfDNA was extracted by modified phenol-chloroform (MPC) method (A) and QIAamp MinElute Virus Spin kit (Qiagen kit) (B). In (A), the height of mutant peak is similar to that of wild-type peak, while the height of mutant peak in (B) is much lower than that of wild-type peak. EGFR, epidermal growth factor receptor.

    Techniques Used: Sequencing, Modification, Mutagenesis

    Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of fragment distribution of plasma cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).
    Figure Legend Snippet: Comparison of cfDNA yield extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit).

    Techniques Used: Modification

    Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Amplification plot of cfDNA extracted by modified phenol-chloroform (MPC) method and QIAamp MinElute Virus Spin kit (Qiagen kit) in DxS EGFR mutation test kit. EGFR, epidermal growth factor receptor.

    Techniques Used: Amplification, Modification, Mutagenesis

    19) Product Images from "Fast-Seq: A Simple Method for Rapid and Inexpensive Validation of Packaged Single-Stranded Adeno-Associated Viral Genomes in Academic Settings"

    Article Title: Fast-Seq: A Simple Method for Rapid and Inexpensive Validation of Packaged Single-Stranded Adeno-Associated Viral Genomes in Academic Settings

    Journal: Human Gene Therapy Methods

    doi: 10.1089/hgtb.2019.110

    Overview of Fast-Seq, a Tn5-based packaged ssAAV genome sequencing method. Preparations of rAAV undergo ssDNA extraction, second-strand synthesis to generate dsDNA, tagmentation with adapter-loaded Tn5 transposomes, indexing, QC validation, short-read sequencing on Illumina MiSeq, mapping reads to the transfer vector plasmid used to generate the input rAAV, and sequence validation of the packaged genome. An example packaged genome mutation (*) is shown at position 7. dsAAV, double-stranded AAV; dsDNA, double-stranded DNA; rAAV, recombinant AAV; ssAAV, single-stranded adeno-associated virus; Tn5, transposase.
    Figure Legend Snippet: Overview of Fast-Seq, a Tn5-based packaged ssAAV genome sequencing method. Preparations of rAAV undergo ssDNA extraction, second-strand synthesis to generate dsDNA, tagmentation with adapter-loaded Tn5 transposomes, indexing, QC validation, short-read sequencing on Illumina MiSeq, mapping reads to the transfer vector plasmid used to generate the input rAAV, and sequence validation of the packaged genome. An example packaged genome mutation (*) is shown at position 7. dsAAV, double-stranded AAV; dsDNA, double-stranded DNA; rAAV, recombinant AAV; ssAAV, single-stranded adeno-associated virus; Tn5, transposase.

    Techniques Used: Sequencing, Plasmid Preparation, Mutagenesis, Recombinant

    Related Articles

    Clone Assay:

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    Article Snippet: Viral RNA was extracted from culture supernatant of DENV2 infected C6/36 cells by using QIAamp MinElute Virus Spin Kit (Qiagen) and reverse transcribed to cDNA by using NS1-R primer (5′-CCG CTC GAG AGC TGT GAC CAA GGA GTT GAC CAA A-3′) and SuperScriptTM III First-Strand Synthesis System for RT-PCR (Invitrogen). .. BamHI and Xho I endonuclease sites (underlined) were incorporated into the primer sequences to facilitate subsequent DNA cloning.

    Centrifugation:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: Three methods were applied for the extraction of cfDNA, i.e. MPC method, traditional phenol-chloroform (PC) method, and commercial QIAamp MinElute Virus Spin kit (Qiagen kit). .. Then, the mixture was centrifuged at 16,000 g at room temperature for 15 min (PC method), or transferred into a Phase Lock Gel tube (MaXtract Low Density tube, Qiagen, Hilden, Germany) followed by centrifugation at 16,000 g at room temperature for 5 min (MPC method).

    Amplification:

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    Filtration:

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    Polymerase Chain Reaction:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: The yield of plasma cfDNA extracted by our modified phenol-chloroform (MPC) method from non-small-cell lung cancer (NSCLC) patients was compared with that by QIAamp MinElute Virus Spin kit (Qiagen kit) as control, using the Wilcoxon rank-sum test. .. Both Mutant-enriched PCR (ME-PCR) coupled sequencing and DxS EGFR mutation test kit were used to evaluate the impact of extraction method on EGFR mutation analysis.

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    Real-time Polymerase Chain Reaction:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
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    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
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    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
    Article Snippet: .. The efficiency of nucleic acid extraction was measured by qPCR and the data are presented as fold change in comparison to the QIAGEN QIAamp® MinElute® Virus Spin kit for total nucleic acid extraction. .. Compared to total nucleic acid extraction using the QIAamp® MinElute® Virus Spin kit, RNA extraction using DNase I did not enhance the recovery of the two single-stranded RNA viruses, however, the RNase A digestion step enriched DNA extraction for double-stranded nucleic acids (both DNA and RNA; Fig. ).

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: Paragraph title: Parvovirus replication and production: real-time qPCR and plaque assay ... Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.

    Incubation:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: Three methods were applied for the extraction of cfDNA, i.e. MPC method, traditional phenol-chloroform (PC) method, and commercial QIAamp MinElute Virus Spin kit (Qiagen kit). .. For PC/MPC method, 1 mL of plasma was mixed with 50 µL of 25% sodium lauryl sulfate (SDS) and 30 µL of 20 mg/mL proteinase K (Qiagen, Hilden, Germany) and incubated at 55℃ for 16 h. After digestion, equal volume of water-saturated phenol was added into the sample and mixed.

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
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    Modification:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
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    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
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    Hybridization:

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    Ligation:

    Article Title: Maternal CD4+ T cells protect against severe congenital cytomegalovirus disease in a novel nonhuman primate model of placental cytomegalovirus transmission
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    Infection:

    Article Title: Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle
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    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: After 24 h, cells were infected with H-1PV at a MOI of 0.01 pfu/cell, in the presence or absence of VPA (1 mM). .. Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.

    Sequencing:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: The yield of plasma cfDNA extracted by our modified phenol-chloroform (MPC) method from non-small-cell lung cancer (NSCLC) patients was compared with that by QIAamp MinElute Virus Spin kit (Qiagen kit) as control, using the Wilcoxon rank-sum test. .. Both Mutant-enriched PCR (ME-PCR) coupled sequencing and DxS EGFR mutation test kit were used to evaluate the impact of extraction method on EGFR mutation analysis.

    Article Title: Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for In Vivo Nonhuman Primate Studies
    Article Snippet: Viral RNA was isolated using the QIAamp MinElute virus spin kit (Qiagen). .. Conditions for the reverse transcription-PCR were as follows: 50 ° C for 60 min; 94 ° C for 2 min; 2 cycles of 94 ° C for 15 s, 60 ° C for 1 min, and 68 ° C for 4 min; 2 cycles of 94 ° C for 15 s, 58 ° C for 1 min, and 68 ° C for 4 min; 40 cycles of 94 ° C for 15 s, 55 ° C for 1 min, and 68 ° C for 4 min; and 68 ° C for 10 min. Each amplicon covered approximately 2.5 kb, and together they spanned the entire viral genome coding sequence.

    Article Title: Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle
    Article Snippet: Viral RNA was extracted from culture supernatant of DENV2 infected C6/36 cells by using QIAamp MinElute Virus Spin Kit (Qiagen) and reverse transcribed to cDNA by using NS1-R primer (5′-CCG CTC GAG AGC TGT GAC CAA GGA GTT GAC CAA A-3′) and SuperScriptTM III First-Strand Synthesis System for RT-PCR (Invitrogen). .. The cDNA was used as a template for amplification of full-length NS1-coding sequence by Platinum® Pfx DNA polymerase (Invitrogen).

    Article Title: Maternal CD4+ T cells protect against severe congenital cytomegalovirus disease in a novel nonhuman primate model of placental cytomegalovirus transmission
    Article Snippet: Paragraph title: rhCMV Deep Sequencing from Plasma. ... Viral DNA was then isolated using the QIAamp MinElute virus spin kit (Qiagen) according to the manufacturer’s instructions, but omitting carrier RNA.

    Binding Assay:

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
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    Cellular Antioxidant Activity Assay:

    Article Title: Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle
    Article Snippet: .. Viral RNA was extracted from culture supernatant of DENV2 infected C6/36 cells by using QIAamp MinElute Virus Spin Kit (Qiagen) and reverse transcribed to cDNA by using NS1-R primer (5′-CCG CTC GAG AGC TGT GAC CAA GGA GTT GAC CAA A-3′) and SuperScriptTM III First-Strand Synthesis System for RT-PCR (Invitrogen). .. The cDNA was used as a template for amplification of full-length NS1-coding sequence by Platinum® Pfx DNA polymerase (Invitrogen).

    DNA Extraction:

    Article Title: A pre-S gene chip to detect pre-S deletions in hepatitis B virus large surface antigen as a predictive marker for hepatoma risk in chronic hepatitis B virus carriers
    Article Snippet: .. A DNA extraction kit (QIAamp MinElute Virus Spin kit; Qiagen Inc., Valencia, CA) was used to extract the virus DNA in serum. .. Most of the common chemicals used for chip hybridization were purchased from Sigma-Aldrich Co., St Louis, MO.

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: Tumor tissue is the most optimal sample type for genomic DNA extraction for EGFR mutation analysis. .. Since QIAamp MinElute Virus Spin kit, in comparison with several other commercially available kits, has been reported to yield the highest amount of cfDNA, we compared our MPC method to Qiagen kit method on cfDNA extraction yield and the impact of different extraction method on EGFR mutation analysis.

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
    Article Snippet: RNA extraction using the QIAGEN RNeasy® Mini kit was performed with an additional DNase I digestion step after binding the nucleic acid to the column and performing an initial wash. DNA extraction using the Wako DNA Extraction® kit was performed with an RNase A digestion step during the initial Proteinase K digestion. .. The efficiency of nucleic acid extraction was measured by qPCR and the data are presented as fold change in comparison to the QIAGEN QIAamp® MinElute® Virus Spin kit for total nucleic acid extraction.

    In Vivo:

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. To assess virus replication in vivo , 50 µl of the cell lysate obtained from tumour xenografts were digested with 50 U/ml of benzonase® Nuclease and subjected to virus DNA purification as described above.

    Plaque Assay:

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: Paragraph title: Parvovirus replication and production: real-time qPCR and plaque assay ... Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.

    Magnetic Beads:

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
    Article Snippet: Extraction methods comprising silica membrane columns, magnetic beads, and nucleic acid precipitation were compared against each other. .. To facilitate the comparison, the QIAGEN QIAamp® MinElute® Virus Spin kit was used as the baseline, as the use of QIAGEN silica membrane-based nucleic acid extraction kits had been previously documented in viral adventitious agent testing.

    Mutagenesis:

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: The yield of plasma cfDNA extracted by our modified phenol-chloroform (MPC) method from non-small-cell lung cancer (NSCLC) patients was compared with that by QIAamp MinElute Virus Spin kit (Qiagen kit) as control, using the Wilcoxon rank-sum test. .. Both Mutant-enriched PCR (ME-PCR) coupled sequencing and DxS EGFR mutation test kit were used to evaluate the impact of extraction method on EGFR mutation analysis.

    Article Title: A Modified Extraction Method of Circulating Free DNA for Epidermal Growth Factor Receptor Mutation Analysis
    Article Snippet: .. Since QIAamp MinElute Virus Spin kit, in comparison with several other commercially available kits, has been reported to yield the highest amount of cfDNA, we compared our MPC method to Qiagen kit method on cfDNA extraction yield and the impact of different extraction method on EGFR mutation analysis. .. Subjects and samples A total of 25 patients with advanced lung adenocarcinoma were recruited from No.3 People's Hospital, School of Medicine, Shanghai Jiaotong University, China between March and September 2009.

    Isolation:

    Article Title: Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for In Vivo Nonhuman Primate Studies
    Article Snippet: .. Viral RNA was isolated using the QIAamp MinElute virus spin kit (Qiagen). .. RNA was then reverse transcribed and amplified in four overlapping amplicons using SIV-specific primers and the Superscript III one-step RT-PCR system with Platinum high-fidelity Taq (Invitrogen).

    Article Title: Maternal CD4+ T cells protect against severe congenital cytomegalovirus disease in a novel nonhuman primate model of placental cytomegalovirus transmission
    Article Snippet: .. Viral DNA was then isolated using the QIAamp MinElute virus spin kit (Qiagen) according to the manufacturer’s instructions, but omitting carrier RNA. .. Approximately 1 ng of DNA was then subjected to simultaneous fragmentation and adaptor ligation with the Nextera XT DNA sample preparation kit (Illumina) to generate deep-sequencing libraries.

    Purification:

    Article Title: Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for In Vivo Nonhuman Primate Studies
    Article Snippet: Viral RNA was isolated using the QIAamp MinElute virus spin kit (Qiagen). .. PCR products were gel purified and quantitated using the Qubit double-stranded DNA (dsDNA) HS assay kit (Invitrogen).

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: .. Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. Quantification of viral DNA was carried out by real-time qPCR, as previously described (El-Andaloussi et al, ).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for In Vivo Nonhuman Primate Studies
    Article Snippet: Viral RNA was isolated using the QIAamp MinElute virus spin kit (Qiagen). .. RNA was then reverse transcribed and amplified in four overlapping amplicons using SIV-specific primers and the Superscript III one-step RT-PCR system with Platinum high-fidelity Taq (Invitrogen).

    Article Title: Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle
    Article Snippet: .. Viral RNA was extracted from culture supernatant of DENV2 infected C6/36 cells by using QIAamp MinElute Virus Spin Kit (Qiagen) and reverse transcribed to cDNA by using NS1-R primer (5′-CCG CTC GAG AGC TGT GAC CAA GGA GTT GAC CAA A-3′) and SuperScriptTM III First-Strand Synthesis System for RT-PCR (Invitrogen). .. The cDNA was used as a template for amplification of full-length NS1-coding sequence by Platinum® Pfx DNA polymerase (Invitrogen).

    Chromatin Immunoprecipitation:

    Article Title: A pre-S gene chip to detect pre-S deletions in hepatitis B virus large surface antigen as a predictive marker for hepatoma risk in chronic hepatitis B virus carriers
    Article Snippet: To test the efficacy of the chip, another 20 serum samples from chronic HBV carriers were obtained from the Center for Disease Control, Taipei, Taiwan, and analyzed using the chip. .. A DNA extraction kit (QIAamp MinElute Virus Spin kit; Qiagen Inc., Valencia, CA) was used to extract the virus DNA in serum.

    RNA Extraction:

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals
    Article Snippet: RNA extraction using the QIAGEN RNeasy® Mini kit was performed with an additional DNase I digestion step after binding the nucleic acid to the column and performing an initial wash. DNA extraction using the Wako DNA Extraction® kit was performed with an RNase A digestion step during the initial Proteinase K digestion. .. The efficiency of nucleic acid extraction was measured by qPCR and the data are presented as fold change in comparison to the QIAGEN QIAamp® MinElute® Virus Spin kit for total nucleic acid extraction.

    Recombinant:

    Article Title: Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle
    Article Snippet: Paragraph title: Preparation of recombinant NS1 (rNS1) ... Viral RNA was extracted from culture supernatant of DENV2 infected C6/36 cells by using QIAamp MinElute Virus Spin Kit (Qiagen) and reverse transcribed to cDNA by using NS1-R primer (5′-CCG CTC GAG AGC TGT GAC CAA GGA GTT GAC CAA A-3′) and SuperScriptTM III First-Strand Synthesis System for RT-PCR (Invitrogen).

    Sample Prep:

    Article Title: Maternal CD4+ T cells protect against severe congenital cytomegalovirus disease in a novel nonhuman primate model of placental cytomegalovirus transmission
    Article Snippet: Viral DNA was then isolated using the QIAamp MinElute virus spin kit (Qiagen) according to the manufacturer’s instructions, but omitting carrier RNA. .. Approximately 1 ng of DNA was then subjected to simultaneous fragmentation and adaptor ligation with the Nextera XT DNA sample preparation kit (Illumina) to generate deep-sequencing libraries.

    DNA Purification:

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas
    Article Snippet: Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. To assess virus replication in vivo , 50 µl of the cell lysate obtained from tumour xenografts were digested with 50 U/ml of benzonase® Nuclease and subjected to virus DNA purification as described above.

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    Qiagen qiaamp minelute virus spin kit
    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using <t>QIAamp®</t> <t>MinElute®</t> Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)
    Qiaamp Minelute Virus Spin Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Journal: NPJ Vaccines

    Article Title: Selection and evaluation of an efficient method for the recovery of viral nucleic acids from complex biologicals

    doi: 10.1038/s41541-018-0067-3

    Figure Lengend Snippet: Efficiency of separate extraction of DNA and RNA when compared to total nucleic acid extraction using QIAamp® MinElute® Virus Spin kit. DNA and RNA were extracted separately and compared to the total nucleic acid extracted using the QIAamp® MinElute® Virus Spin kit, using qPCR. RNA extraction using the Qiagen RNeasy® Mini kit did not enhance the detection of the two single-stranded RNA viruses (FeLV and RSV), whereas the RNase A digestion added to the Wako DNA Extractor® kit greatly enriched for double-stranded nucleic acid (EBV and Reo3)

    Article Snippet: The data are presented as fold change in comparison to the QIAGEN QIAamp® MinElute® Virus Spin kit.

    Techniques: Real-time Polymerase Chain Reaction, RNA Extraction

    H-1PV/VPA co-treatment leads to complete remission of established HeLa tumours Source data is available for this figure in the Supporting Information. Analysis of tumour growth. Animals ( n = 8 per group) were treated with either PBS (control), VPA or a combination of both agents as described in Materials and Methods Section. Other animal groups are presented in Supporting Information Fig S7 . The data shown represent average tumour volumes with standard deviation bars. Graphical summary of Kaplan–Meier survival analysis of tumour-bearing nude rats treated as indicated. Animals were sacrificed when the tumour reached the maximum tolerable size of 4000 mm 3 according to law. The difference between H-1PV/VPA co-treatment and treatment with H-1PV alone was statistically significant ( p = 0.0002 as calculated with the two-sided log-rank test and adjusted for multiple testing with the Bonferroni method). Other results from the statistical analysis are shown in Supporting Information Table S2. H-1PV/VPA co-treatment induces DNA damage and apoptosis in vivo . Two rats of each treatment group were sacrificed on day 19. Tumours were fixed, sectioned and examined morphologically either by H E staining or by immunochemistry using antibodies against NS1, VP capsid proteins, γ-H2AX (a DNA damage marker), or active cleaved caspase-3 (an apoptosis marker). Scale bar = 50 µm. Higher levels of NS1, VP1 and VP2 viral proteins in tumour samples from VPA co-treated animals. Tumour-bearing animals ( n = 3) treated with H-1PV alone (total virus dose of 3.6 × 10 9 Vg/animal corresponding to 1.5 × 10 8 pfu/animals subdivided into two subdoses given at days 0 and 7) or co-treated with H-1PV and VPA (100 mg/kg) were sacrificed at day 14. Complete tumours were resected, homogenised and lysed. 20 µg of total cellular extracts were analysed by SDS–PAGE for the presence of the viral proteins. Actin was used as a loading control. VPA-enhanced virus multiplication in vivo . Viral particles were recovered from aliquots of tumour extracts from animals treated as in panel D, purified by using the QIAamp MinElute Virus Spin Kit (Qiagen) and quantified by real time qPCR. Columns represent the average amounts of the virus recovered from the tumours of three animals with standard deviation bars. Numbers on top of the columns indicate the fold increase in virus titers in the presence versus absence of VPA.

    Journal: EMBO Molecular Medicine

    Article Title: Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas

    doi: 10.1002/emmm.201302796

    Figure Lengend Snippet: H-1PV/VPA co-treatment leads to complete remission of established HeLa tumours Source data is available for this figure in the Supporting Information. Analysis of tumour growth. Animals ( n = 8 per group) were treated with either PBS (control), VPA or a combination of both agents as described in Materials and Methods Section. Other animal groups are presented in Supporting Information Fig S7 . The data shown represent average tumour volumes with standard deviation bars. Graphical summary of Kaplan–Meier survival analysis of tumour-bearing nude rats treated as indicated. Animals were sacrificed when the tumour reached the maximum tolerable size of 4000 mm 3 according to law. The difference between H-1PV/VPA co-treatment and treatment with H-1PV alone was statistically significant ( p = 0.0002 as calculated with the two-sided log-rank test and adjusted for multiple testing with the Bonferroni method). Other results from the statistical analysis are shown in Supporting Information Table S2. H-1PV/VPA co-treatment induces DNA damage and apoptosis in vivo . Two rats of each treatment group were sacrificed on day 19. Tumours were fixed, sectioned and examined morphologically either by H E staining or by immunochemistry using antibodies against NS1, VP capsid proteins, γ-H2AX (a DNA damage marker), or active cleaved caspase-3 (an apoptosis marker). Scale bar = 50 µm. Higher levels of NS1, VP1 and VP2 viral proteins in tumour samples from VPA co-treated animals. Tumour-bearing animals ( n = 3) treated with H-1PV alone (total virus dose of 3.6 × 10 9 Vg/animal corresponding to 1.5 × 10 8 pfu/animals subdivided into two subdoses given at days 0 and 7) or co-treated with H-1PV and VPA (100 mg/kg) were sacrificed at day 14. Complete tumours were resected, homogenised and lysed. 20 µg of total cellular extracts were analysed by SDS–PAGE for the presence of the viral proteins. Actin was used as a loading control. VPA-enhanced virus multiplication in vivo . Viral particles were recovered from aliquots of tumour extracts from animals treated as in panel D, purified by using the QIAamp MinElute Virus Spin Kit (Qiagen) and quantified by real time qPCR. Columns represent the average amounts of the virus recovered from the tumours of three animals with standard deviation bars. Numbers on top of the columns indicate the fold increase in virus titers in the presence versus absence of VPA.

    Article Snippet: Viral genomic DNA was purified from cell lysates by using the QiaAmp MinElute virus kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.

    Techniques: Standard Deviation, In Vivo, Staining, Marker, SDS Page, Purification, Real-time Polymerase Chain Reaction