qiaamp viral rna mini kit  (Qiagen)


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    Name:
    QIAamp Viral RNA Mini Kit
    Description:
    For isolation of viral RNA from cell free body fluids Kit contents Qiagen QIAamp Viral RNA Mini Kit 50 preps 140L Sample 50L Elution Volume Liquid Media Sample Viral RNA Purification Silica Technology Spin Column Format Manual Processing 90 Yield 20 to 40 min Time Run Ideal for PCR qPCR Real time PCR For Isolation of Viral RNA from Cell free Body Fluids Includes 50 QIAamp Mini Spin Columns Carrier RNA 2mL Collection Tubes RNase free Buffers Benefits Rapid isolation of high quality ready to use RNA No organic extraction or alcohol precipitation Consistent high yields Complete removal of contaminants and inhibitors
    Catalog Number:
    52904
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    QIAamp Viral RNA Mini Kit
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    Qiagen qiaamp viral rna mini kit
    QIAamp Viral RNA Mini Kit
    For isolation of viral RNA from cell free body fluids Kit contents Qiagen QIAamp Viral RNA Mini Kit 50 preps 140L Sample 50L Elution Volume Liquid Media Sample Viral RNA Purification Silica Technology Spin Column Format Manual Processing 90 Yield 20 to 40 min Time Run Ideal for PCR qPCR Real time PCR For Isolation of Viral RNA from Cell free Body Fluids Includes 50 QIAamp Mini Spin Columns Carrier RNA 2mL Collection Tubes RNase free Buffers Benefits Rapid isolation of high quality ready to use RNA No organic extraction or alcohol precipitation Consistent high yields Complete removal of contaminants and inhibitors
    https://www.bioz.com/result/qiaamp viral rna mini kit/product/Qiagen
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    qiaamp viral rna mini kit - by Bioz Stars, 2021-06
    97/100 stars

    Images

    1) Product Images from "Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology"

    Article Title: Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S191784

    Quantitative analysis of the viral gene of rotavirus adsorbed onto the antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to RT-reaction. The resultant cDNA was analyzed by real-time PCR using primers for the rotavirus VP7 gene as described in Materials and methods. The value of the TL sample was taken as 100%. Abbreviations: MNBs, magnetic nanobeads; RT, reverse transcription.
    Figure Legend Snippet: Quantitative analysis of the viral gene of rotavirus adsorbed onto the antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to RT-reaction. The resultant cDNA was analyzed by real-time PCR using primers for the rotavirus VP7 gene as described in Materials and methods. The value of the TL sample was taken as 100%. Abbreviations: MNBs, magnetic nanobeads; RT, reverse transcription.

    Techniques Used: Infection, Incubation, Magnetic Beads, Real-time Polymerase Chain Reaction

    Detection of viral RNA of rotavirus adsorbed onto antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction. Rotavirus viral protein 7 (VP7) gene (552 bp) in the cDNA was amplified by PCR as described in Materials and methods. PCR products were analyzed by agarose gel electrophoresis (1.2% gel). The identity of the amplified products was confirmed by DNA sequencing. The left-hand lane is size marker (M), which includes DNA of 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,200, and 1,500 bp. The position of the 552 bp band for VP7 is indicated by an arrow. The NC comprised a water sample (no rotavirus) that was subjected to RT-PCR. Abbreviations: MNBs, magnetic nanobeads; NC, negative control; RT, reverse transcription.
    Figure Legend Snippet: Detection of viral RNA of rotavirus adsorbed onto antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction. Rotavirus viral protein 7 (VP7) gene (552 bp) in the cDNA was amplified by PCR as described in Materials and methods. PCR products were analyzed by agarose gel electrophoresis (1.2% gel). The identity of the amplified products was confirmed by DNA sequencing. The left-hand lane is size marker (M), which includes DNA of 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,200, and 1,500 bp. The position of the 552 bp band for VP7 is indicated by an arrow. The NC comprised a water sample (no rotavirus) that was subjected to RT-PCR. Abbreviations: MNBs, magnetic nanobeads; NC, negative control; RT, reverse transcription.

    Techniques Used: Infection, Incubation, Magnetic Beads, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, DNA Sequencing, Marker, Reverse Transcription Polymerase Chain Reaction, Negative Control

    2) Product Images from "Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5"

    Article Title: Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5

    Journal: Journal of Virology

    doi: 10.1128/JVI.01832-15

    Effects of V mutants on viral RNA synthesis. Truncated V proteins inhibited PIV5 RNA replication. BSR T7 cells were transfected as described in Materials and Methods. Encapsidated PIV5 minigenome RNA was purified with NP antibody, and a primer that binds to antigenomic RNA in the trailer region was added to the reverse transcription reaction mixture. Then, the same amount of RNA from the reverse transcription reaction was added to the PCR mixture as the template, and a pair of primers that anneal to the C terminus of Renilla luciferase and the PIV5 trailer region, respectively, was also added to the PCR mixture. The expected PCR products are indicated by MG and were approximately 250 bp. DNA ladder, a 100-bp DNA size marker (the bands of the ladder at 200 bp are indicated). (A) Detection of minigenome RNA using RT-PCR. Purified RNA without RT was used in the PCR mixture to demonstrate the lack of plasmid DNA contamination, and the corresponding RNA with RT was used in the PCR mixture. All samples were amplified for 30 cycles. (B) Detection of minigenome RNA using RT-PCR and different numbers of PCR cycles. Purified RNAs were reverse transcribed and then amplified for 15, 20, 25, and 30 cycles. No MG, cells transfected with all minigenome components except the plasmid carrying the minigenome; Pos Control, a minigenome system without plasmid carrying V or the V mutants; instead, the same expression plasmid containing enhanced GFP was used to maintain the total amount of transfected plasmid DNA constant. This gel is representative of those from three experiments. (C) Analysis of PCR products. The intensity of the PCR products was quantified using ImageQuant software. The intensity of cells transfected with all minigenome components except the plasmid carrying the minigenome was set equal to 0 in PCRs with 25 and 30 cycles; the intensity of the positive control was set to 100 in PCRs with 25 and 30 cycles; the relative percentage of the V, V120, or V21-222 PCR band intensity is the ratio of the V, V120, or V21-222 band intensity to that of the positive control in 25 and 30 cycles. The average was calculated from three independent experiments. Error bars represent standards deviation of the means. *, P
    Figure Legend Snippet: Effects of V mutants on viral RNA synthesis. Truncated V proteins inhibited PIV5 RNA replication. BSR T7 cells were transfected as described in Materials and Methods. Encapsidated PIV5 minigenome RNA was purified with NP antibody, and a primer that binds to antigenomic RNA in the trailer region was added to the reverse transcription reaction mixture. Then, the same amount of RNA from the reverse transcription reaction was added to the PCR mixture as the template, and a pair of primers that anneal to the C terminus of Renilla luciferase and the PIV5 trailer region, respectively, was also added to the PCR mixture. The expected PCR products are indicated by MG and were approximately 250 bp. DNA ladder, a 100-bp DNA size marker (the bands of the ladder at 200 bp are indicated). (A) Detection of minigenome RNA using RT-PCR. Purified RNA without RT was used in the PCR mixture to demonstrate the lack of plasmid DNA contamination, and the corresponding RNA with RT was used in the PCR mixture. All samples were amplified for 30 cycles. (B) Detection of minigenome RNA using RT-PCR and different numbers of PCR cycles. Purified RNAs were reverse transcribed and then amplified for 15, 20, 25, and 30 cycles. No MG, cells transfected with all minigenome components except the plasmid carrying the minigenome; Pos Control, a minigenome system without plasmid carrying V or the V mutants; instead, the same expression plasmid containing enhanced GFP was used to maintain the total amount of transfected plasmid DNA constant. This gel is representative of those from three experiments. (C) Analysis of PCR products. The intensity of the PCR products was quantified using ImageQuant software. The intensity of cells transfected with all minigenome components except the plasmid carrying the minigenome was set equal to 0 in PCRs with 25 and 30 cycles; the intensity of the positive control was set to 100 in PCRs with 25 and 30 cycles; the relative percentage of the V, V120, or V21-222 PCR band intensity is the ratio of the V, V120, or V21-222 band intensity to that of the positive control in 25 and 30 cycles. The average was calculated from three independent experiments. Error bars represent standards deviation of the means. *, P

    Techniques Used: Transfection, Purification, Polymerase Chain Reaction, Luciferase, Marker, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Amplification, Expressing, Software, Positive Control

    3) Product Images from "Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin, et al. Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin"

    Article Title: Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin, et al. Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin

    Journal: Transfusion

    doi: 10.1111/trf.16014

    Characterization of untreated, S/D‐treated and S/D and protease–treated HEV. Buoyant densities of, A, wild‐type HEV from human plasma, B, rHEV, and C, wild‐type HEV from porcine feces without any further treatment (untreated), following S/D treatment (S/D treated), or after S/D and protease (pronase E) treatment. Virus was resolved in isopycnic gradients and RNA was detected by RT qPCR. LE particles have a density of 1.06 to 1.10 g/cm 3 , virions of IM band at 1.15 to 1.17 g/cm 3 , whereas NLE virions have a density of 1.19 to 1.22 g/cm 3
    Figure Legend Snippet: Characterization of untreated, S/D‐treated and S/D and protease–treated HEV. Buoyant densities of, A, wild‐type HEV from human plasma, B, rHEV, and C, wild‐type HEV from porcine feces without any further treatment (untreated), following S/D treatment (S/D treated), or after S/D and protease (pronase E) treatment. Virus was resolved in isopycnic gradients and RNA was detected by RT qPCR. LE particles have a density of 1.06 to 1.10 g/cm 3 , virions of IM band at 1.15 to 1.17 g/cm 3 , whereas NLE virions have a density of 1.19 to 1.22 g/cm 3

    Techniques Used: Quantitative RT-PCR

    Related Articles

    Incubation:

    Article Title: Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5
    Article Snippet: Cells were lysed with MCLB lysis buffer, and clarified lysate supernatant was obtained after centrifugation at 12,000 rpm for 10 min. .. The lysate was incubated with anti-NP214 antibody and recombinant protein G-Sepharose 4B beads at 4°C for 3 h. The beads were washed with PBS 3 times, and NP-encapsidated RNA was processed with a mini-viral RNA kit (Qiagen) and then treated with RNase-free DNase (Qiagen) for 3 h at 37°C to eliminate potential plasmid DNA contamination. .. The RNA was purified and dissolved in 30 μl RNase-free water using a mini-RNeasy kit (Qiagen).

    Recombinant:

    Article Title: Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5
    Article Snippet: Cells were lysed with MCLB lysis buffer, and clarified lysate supernatant was obtained after centrifugation at 12,000 rpm for 10 min. .. The lysate was incubated with anti-NP214 antibody and recombinant protein G-Sepharose 4B beads at 4°C for 3 h. The beads were washed with PBS 3 times, and NP-encapsidated RNA was processed with a mini-viral RNA kit (Qiagen) and then treated with RNase-free DNase (Qiagen) for 3 h at 37°C to eliminate potential plasmid DNA contamination. .. The RNA was purified and dissolved in 30 μl RNase-free water using a mini-RNeasy kit (Qiagen).

    Plasmid Preparation:

    Article Title: Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5
    Article Snippet: Cells were lysed with MCLB lysis buffer, and clarified lysate supernatant was obtained after centrifugation at 12,000 rpm for 10 min. .. The lysate was incubated with anti-NP214 antibody and recombinant protein G-Sepharose 4B beads at 4°C for 3 h. The beads were washed with PBS 3 times, and NP-encapsidated RNA was processed with a mini-viral RNA kit (Qiagen) and then treated with RNase-free DNase (Qiagen) for 3 h at 37°C to eliminate potential plasmid DNA contamination. .. The RNA was purified and dissolved in 30 μl RNase-free water using a mini-RNeasy kit (Qiagen).

    other:

    Article Title: Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
    Article Snippet: Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction.

    Article Title: First Report of the Presence of Hepatitis E Virus in Scottish-Harvested Shellfish Purchased at Retail Level
    Article Snippet: Viral RNA was extracted from shellfish supernatants using QIamp Viral RNA Mini Kit (Qiagen, Hilden, Germany).

    Cell Culture:

    Article Title: Apoptotic and Early Innate Immune Responses to PB1-F2 Protein of Influenza A Viruses Belonging to Different Subtypes in Human Lung Epithelial A549 Cells
    Article Snippet: .. The cell culture supernatant was used to extract viral RNA with QIAamp Viral RNA Mini Kit (Qiagen, Germany). .. Reverse transcription was carried out using Superscript II RT-PCR kit (Invitrogen, Carlsbad, CA, USA) and Uni12 primer [ ].

    Isolation:

    Article Title: Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin, et al. Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin
    Article Snippet: For analysis of HEV RNA content, two sequential fractions were pooled before RNA extraction. .. 2.4 Detection of HEV RNAHEV RNA was extracted from volumes of 120 μL with a viral RNA isolation kit (QIAamp; Qiagen, Hilden, Germany). .. Samples were eluted in 80 μL of buffer provided with the kit and reverse transcription quantitative polymerase chain reaction (RT qPCR) was performed on a RT qPCR system (ABI 7900HT Real‐Time PCR; Applied Biosystems, Life Technologies, Carlsbad, California) with an optimized HEV RT qPCR assay, as previously reported.

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    Qiagen qiaamp viral rna mini kit
    Quantitative analysis of the viral gene of rotavirus adsorbed onto the antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic <t>RNA</t> was subsequently extracted from the above fractions using a <t>QIAamp</t> Viral RNA mini kit and subjected to RT-reaction. The resultant cDNA was analyzed by real-time PCR using primers for the rotavirus VP7 gene as described in Materials and methods. The value of the TL sample was taken as 100%. Abbreviations: MNBs, magnetic nanobeads; RT, reverse transcription.
    Qiaamp Viral Rna Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qiaamp viral rna mini kit/product/Qiagen
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    qiaamp viral rna mini kit - by Bioz Stars, 2021-06
    97/100 stars
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    Quantitative analysis of the viral gene of rotavirus adsorbed onto the antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to RT-reaction. The resultant cDNA was analyzed by real-time PCR using primers for the rotavirus VP7 gene as described in Materials and methods. The value of the TL sample was taken as 100%. Abbreviations: MNBs, magnetic nanobeads; RT, reverse transcription.

    Journal: International Journal of Nanomedicine

    Article Title: Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology

    doi: 10.2147/IJN.S191784

    Figure Lengend Snippet: Quantitative analysis of the viral gene of rotavirus adsorbed onto the antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to RT-reaction. The resultant cDNA was analyzed by real-time PCR using primers for the rotavirus VP7 gene as described in Materials and methods. The value of the TL sample was taken as 100%. Abbreviations: MNBs, magnetic nanobeads; RT, reverse transcription.

    Article Snippet: Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction.

    Techniques: Infection, Incubation, Magnetic Beads, Real-time Polymerase Chain Reaction

    Detection of viral RNA of rotavirus adsorbed onto antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction. Rotavirus viral protein 7 (VP7) gene (552 bp) in the cDNA was amplified by PCR as described in Materials and methods. PCR products were analyzed by agarose gel electrophoresis (1.2% gel). The identity of the amplified products was confirmed by DNA sequencing. The left-hand lane is size marker (M), which includes DNA of 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,200, and 1,500 bp. The position of the 552 bp band for VP7 is indicated by an arrow. The NC comprised a water sample (no rotavirus) that was subjected to RT-PCR. Abbreviations: MNBs, magnetic nanobeads; NC, negative control; RT, reverse transcription.

    Journal: International Journal of Nanomedicine

    Article Title: Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology

    doi: 10.2147/IJN.S191784

    Figure Lengend Snippet: Detection of viral RNA of rotavirus adsorbed onto antibody-integrated MNBs. Notes: Rotavirus-infected cell lysate (10 µL) was diluted with PBS (500 µL) and then incubated with antibody-integrated magnetic beads. After incubation, the following fractions were obtained: 1) diluted rotavirus sample before incubation with the beads (BF), 2) bead fraction after incubation with anti-rotavirus antibody-integrated MNBs (RV-BD), 3) bead fraction after incubation with anti-dengue virus antibody-integrated MNBs (DV-BD), 4) supernatant fraction after incubation with the anti-rotavirus antibody-integrated MNBs (RV-SP), 5) supernatant fraction after incubation with the anti-dengue virus antibody-integrated MNBs (DV-SP), and 6) total sample containing the same quantity of rotavirus as in 10 µL of rotavirus-infected cell lysate (total fraction, TL). Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction. Rotavirus viral protein 7 (VP7) gene (552 bp) in the cDNA was amplified by PCR as described in Materials and methods. PCR products were analyzed by agarose gel electrophoresis (1.2% gel). The identity of the amplified products was confirmed by DNA sequencing. The left-hand lane is size marker (M), which includes DNA of 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,200, and 1,500 bp. The position of the 552 bp band for VP7 is indicated by an arrow. The NC comprised a water sample (no rotavirus) that was subjected to RT-PCR. Abbreviations: MNBs, magnetic nanobeads; NC, negative control; RT, reverse transcription.

    Article Snippet: Viral genomic RNA was subsequently extracted from the above fractions using a QIAamp Viral RNA mini kit and subjected to a RT-reaction.

    Techniques: Infection, Incubation, Magnetic Beads, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, DNA Sequencing, Marker, Reverse Transcription Polymerase Chain Reaction, Negative Control

    Effects of V mutants on viral RNA synthesis. Truncated V proteins inhibited PIV5 RNA replication. BSR T7 cells were transfected as described in Materials and Methods. Encapsidated PIV5 minigenome RNA was purified with NP antibody, and a primer that binds to antigenomic RNA in the trailer region was added to the reverse transcription reaction mixture. Then, the same amount of RNA from the reverse transcription reaction was added to the PCR mixture as the template, and a pair of primers that anneal to the C terminus of Renilla luciferase and the PIV5 trailer region, respectively, was also added to the PCR mixture. The expected PCR products are indicated by MG and were approximately 250 bp. DNA ladder, a 100-bp DNA size marker (the bands of the ladder at 200 bp are indicated). (A) Detection of minigenome RNA using RT-PCR. Purified RNA without RT was used in the PCR mixture to demonstrate the lack of plasmid DNA contamination, and the corresponding RNA with RT was used in the PCR mixture. All samples were amplified for 30 cycles. (B) Detection of minigenome RNA using RT-PCR and different numbers of PCR cycles. Purified RNAs were reverse transcribed and then amplified for 15, 20, 25, and 30 cycles. No MG, cells transfected with all minigenome components except the plasmid carrying the minigenome; Pos Control, a minigenome system without plasmid carrying V or the V mutants; instead, the same expression plasmid containing enhanced GFP was used to maintain the total amount of transfected plasmid DNA constant. This gel is representative of those from three experiments. (C) Analysis of PCR products. The intensity of the PCR products was quantified using ImageQuant software. The intensity of cells transfected with all minigenome components except the plasmid carrying the minigenome was set equal to 0 in PCRs with 25 and 30 cycles; the intensity of the positive control was set to 100 in PCRs with 25 and 30 cycles; the relative percentage of the V, V120, or V21-222 PCR band intensity is the ratio of the V, V120, or V21-222 band intensity to that of the positive control in 25 and 30 cycles. The average was calculated from three independent experiments. Error bars represent standards deviation of the means. *, P

    Journal: Journal of Virology

    Article Title: Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5

    doi: 10.1128/JVI.01832-15

    Figure Lengend Snippet: Effects of V mutants on viral RNA synthesis. Truncated V proteins inhibited PIV5 RNA replication. BSR T7 cells were transfected as described in Materials and Methods. Encapsidated PIV5 minigenome RNA was purified with NP antibody, and a primer that binds to antigenomic RNA in the trailer region was added to the reverse transcription reaction mixture. Then, the same amount of RNA from the reverse transcription reaction was added to the PCR mixture as the template, and a pair of primers that anneal to the C terminus of Renilla luciferase and the PIV5 trailer region, respectively, was also added to the PCR mixture. The expected PCR products are indicated by MG and were approximately 250 bp. DNA ladder, a 100-bp DNA size marker (the bands of the ladder at 200 bp are indicated). (A) Detection of minigenome RNA using RT-PCR. Purified RNA without RT was used in the PCR mixture to demonstrate the lack of plasmid DNA contamination, and the corresponding RNA with RT was used in the PCR mixture. All samples were amplified for 30 cycles. (B) Detection of minigenome RNA using RT-PCR and different numbers of PCR cycles. Purified RNAs were reverse transcribed and then amplified for 15, 20, 25, and 30 cycles. No MG, cells transfected with all minigenome components except the plasmid carrying the minigenome; Pos Control, a minigenome system without plasmid carrying V or the V mutants; instead, the same expression plasmid containing enhanced GFP was used to maintain the total amount of transfected plasmid DNA constant. This gel is representative of those from three experiments. (C) Analysis of PCR products. The intensity of the PCR products was quantified using ImageQuant software. The intensity of cells transfected with all minigenome components except the plasmid carrying the minigenome was set equal to 0 in PCRs with 25 and 30 cycles; the intensity of the positive control was set to 100 in PCRs with 25 and 30 cycles; the relative percentage of the V, V120, or V21-222 PCR band intensity is the ratio of the V, V120, or V21-222 band intensity to that of the positive control in 25 and 30 cycles. The average was calculated from three independent experiments. Error bars represent standards deviation of the means. *, P

    Article Snippet: The lysate was incubated with anti-NP214 antibody and recombinant protein G-Sepharose 4B beads at 4°C for 3 h. The beads were washed with PBS 3 times, and NP-encapsidated RNA was processed with a mini-viral RNA kit (Qiagen) and then treated with RNase-free DNase (Qiagen) for 3 h at 37°C to eliminate potential plasmid DNA contamination.

    Techniques: Transfection, Purification, Polymerase Chain Reaction, Luciferase, Marker, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Amplification, Expressing, Software, Positive Control

    Characterization of untreated, S/D‐treated and S/D and protease–treated HEV. Buoyant densities of, A, wild‐type HEV from human plasma, B, rHEV, and C, wild‐type HEV from porcine feces without any further treatment (untreated), following S/D treatment (S/D treated), or after S/D and protease (pronase E) treatment. Virus was resolved in isopycnic gradients and RNA was detected by RT qPCR. LE particles have a density of 1.06 to 1.10 g/cm 3 , virions of IM band at 1.15 to 1.17 g/cm 3 , whereas NLE virions have a density of 1.19 to 1.22 g/cm 3

    Journal: Transfusion

    Article Title: Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin, et al. Antibody‐enhanced hepatitis E virus nanofiltration during the manufacture of human immunoglobulin

    doi: 10.1111/trf.16014

    Figure Lengend Snippet: Characterization of untreated, S/D‐treated and S/D and protease–treated HEV. Buoyant densities of, A, wild‐type HEV from human plasma, B, rHEV, and C, wild‐type HEV from porcine feces without any further treatment (untreated), following S/D treatment (S/D treated), or after S/D and protease (pronase E) treatment. Virus was resolved in isopycnic gradients and RNA was detected by RT qPCR. LE particles have a density of 1.06 to 1.10 g/cm 3 , virions of IM band at 1.15 to 1.17 g/cm 3 , whereas NLE virions have a density of 1.19 to 1.22 g/cm 3

    Article Snippet: 2.4 Detection of HEV RNAHEV RNA was extracted from volumes of 120 μL with a viral RNA isolation kit (QIAamp; Qiagen, Hilden, Germany).

    Techniques: Quantitative RT-PCR