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Promega rq dnase i
The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with <t>DNase</t> I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.
Rq Dnase I, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties"

Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties

Journal: Virology

doi: 10.1016/j.virol.2006.07.013

The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.
Figure Legend Snippet: The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.

Techniques Used: Immunofluorescence, Transfection, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification

2) Product Images from "The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties"

Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties

Journal: Virology

doi: 10.1016/j.virol.2006.07.013

The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.
Figure Legend Snippet: The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.

Techniques Used: Immunofluorescence, Transfection, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification

3) Product Images from "The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties"

Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties

Journal: Virology

doi: 10.1016/j.virol.2006.07.013

The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.
Figure Legend Snippet: The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.

Techniques Used: Immunofluorescence, Transfection, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification

4) Product Images from "Restricted Specificity of Xenopus TFIIIA for Transcription of Somatic 5S rRNA Genes"

Article Title: Restricted Specificity of Xenopus TFIIIA for Transcription of Somatic 5S rRNA Genes

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.24.6.2467-2477.2004

Substitutions at serine-16 do not affect the binding affinity of TFIIIA for either 5S rRNA gene. Wild-type TFIIIA or mutant TFIIIA with serine-16 replaced with alanine (S16A) or glutamic acid (S16E) was used in DNase I footprinting assays with the oocyte- or somatic-type 5S rRNA gene. Each sample contained 2 nM end-labeled, linearized plasmid and 0, 10, 15, 20, 30, 40, or 100 nM protein (oocyte gene) or 0, 10, 20, 30, or 40 nM protein (somatic gene).
Figure Legend Snippet: Substitutions at serine-16 do not affect the binding affinity of TFIIIA for either 5S rRNA gene. Wild-type TFIIIA or mutant TFIIIA with serine-16 replaced with alanine (S16A) or glutamic acid (S16E) was used in DNase I footprinting assays with the oocyte- or somatic-type 5S rRNA gene. Each sample contained 2 nM end-labeled, linearized plasmid and 0, 10, 15, 20, 30, 40, or 100 nM protein (oocyte gene) or 0, 10, 20, 30, or 40 nM protein (somatic gene).

Techniques Used: Binding Assay, Mutagenesis, Footprinting, Labeling, Plasmid Preparation

Related Articles

RNA Extraction:

Article Title: Staufen1-mediated mRNA decay induces Requiem mRNA decay through binding of Staufen1 to the Requiem 3?UTR
Article Snippet: .. To eliminate possible contamination by exogenous or endogenous DNA, the RNA was treated with RQ DNase I (Promega) for 30 min at 37°C before continuing with the RNA extraction protocol. .. The quantity and quality of the isolated RNA were confirmed using UV spectroscopy (Shimadzu, Japan) and electrophoresis on a 1% agarose gel, respectively.

Synthesized:

Article Title: Effects of low-dose rate γ-irradiation combined with simulated microgravity on markers of oxidative stress, DNA methylation potential, and remodeling in the mouse heart
Article Snippet: .. DNA was removed from the RNA with RQ-DNAse I (Promega), and cDNA was synthesized from the RNA using a High Capacity cDNA Reverse Transcription Kit (Life Technologies). .. Steady-state mRNA was assessed by real-time quantitative PCR (RT-qPCR).

Isolation:

Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties
Article Snippet: .. RNA was isolated from each sample by using a QIAamp viral RNA mini-kit and subsequently, treated one more time with 1 unit of RQ DNase I. ..

Purification:

Article Title: Restricted Specificity of Xenopus TFIIIA for Transcription of Somatic 5S rRNA Genes
Article Snippet: .. After digestion of the DNA template with RQ DNase I (Promega), the mRNA was purified by passage through a Microspin S-300 HR column (Amersham). .. Internally radiolabeled 5S rRNA was prepared by runoff transcription of pT75S linearized with Dra I with T7 RNA polymerase ( ).

Incubation:

Article Title: Molecular switch-like regulation enables global subunit coordination in a viral ring ATPase
Article Snippet: .. The sample was incubated for 2 min; 1 U of RQ DNase I (Promega) was added and incubated at room temperature for 10 min. .. The sample was placed on ice until grid preparation for cryo-EM imaging.

Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties
Article Snippet: .. Seventy microliters of each sample suspension were added with 2 μl of RQ DNase I (1 U/μl; Promega) and 3.6 μl of RNase A (10 mg/ml; Sigma), and the mixture was incubated for 1 h at 37 °C in the presence or absence of detergents (1 μl of Triton X-100, and 3.5 μl of 10% SDS). .. After incubation for 1 h at 37 °C, the nucleases were inactivated by the addition of 5 μl of proteinase K (20 mg/ml; Qiagen) and incubation for 30 min at 50 °C.

other:

Article Title: Characterization of Influenza Virus NS1 Protein by Using a Novel Helper-Virus-Free Reverse Genetic System
Article Snippet: This digestion condition was determined using different amounts of RQ DNase I.

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    Promega rq dnase i
    The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with <t>DNase</t> I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.
    Rq Dnase I, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rq dnase i/product/Promega
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    rq dnase i - by Bioz Stars, 2020-05
    93/100 stars
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    The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.

    Journal: Virology

    Article Title: The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties

    doi: 10.1016/j.virol.2006.07.013

    Figure Lengend Snippet: The E protein is essential for PRRSV replication. (A) Immunofluorescence of N protein in P129-ΔE inoculated cells. The ‘passage-1’ virus was prepared as culture supernatant harvested from BHK-21 cells transfected with P129-WT or P129-ΔE clone. Marc-145 cells were inoculated with ‘passage-1’ and fixed at the indicated times post-inoculation, followed by staining with N-specific MAb. (B) Detection of viral RNA in culture supernatants and in Marc-145 cells inoculated with ‘passage-1’ or ‘passage-2’ virus. Total RNA was extracted and treated with DNase I followed by RT-PCR for E gene. (C) Strand-specific detection of viral RNA in cells by RT-PCR. Marc-145 cells were inoculated with ‘passage-1’ P129-WT or ‘passage-1’ P129-ΔE, and total cellular RNA was extracted at 2 days post-inoculation. The RNA was treated by DNase I and RT-PCR was conducted to amplify the region as illustrated in the figure. Numbers in parenthesis indicate the 5′ most nucleotide position in each primer with respect to the PRRSV genome. Expected sizes of amplified products are indicated on the right of the gel.

    Article Snippet: Seventy microliters of each sample suspension were added with 2 μl of RQ DNase I (1 U/μl; Promega) and 3.6 μl of RNase A (10 mg/ml; Sigma), and the mixture was incubated for 1 h at 37 °C in the presence or absence of detergents (1 μl of Triton X-100, and 3.5 μl of 10% SDS).

    Techniques: Immunofluorescence, Transfection, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification

    Substitutions at serine-16 do not affect the binding affinity of TFIIIA for either 5S rRNA gene. Wild-type TFIIIA or mutant TFIIIA with serine-16 replaced with alanine (S16A) or glutamic acid (S16E) was used in DNase I footprinting assays with the oocyte- or somatic-type 5S rRNA gene. Each sample contained 2 nM end-labeled, linearized plasmid and 0, 10, 15, 20, 30, 40, or 100 nM protein (oocyte gene) or 0, 10, 20, 30, or 40 nM protein (somatic gene).

    Journal: Molecular and Cellular Biology

    Article Title: Restricted Specificity of Xenopus TFIIIA for Transcription of Somatic 5S rRNA Genes

    doi: 10.1128/MCB.24.6.2467-2477.2004

    Figure Lengend Snippet: Substitutions at serine-16 do not affect the binding affinity of TFIIIA for either 5S rRNA gene. Wild-type TFIIIA or mutant TFIIIA with serine-16 replaced with alanine (S16A) or glutamic acid (S16E) was used in DNase I footprinting assays with the oocyte- or somatic-type 5S rRNA gene. Each sample contained 2 nM end-labeled, linearized plasmid and 0, 10, 15, 20, 30, 40, or 100 nM protein (oocyte gene) or 0, 10, 20, 30, or 40 nM protein (somatic gene).

    Article Snippet: After digestion of the DNA template with RQ DNase I (Promega), the mRNA was purified by passage through a Microspin S-300 HR column (Amersham).

    Techniques: Binding Assay, Mutagenesis, Footprinting, Labeling, Plasmid Preparation