Structured Review

GE Healthcare rnase v1
Analysis of the complex formed between HTNV N protein and deletion RNAs by UV cross-linking analysis. The concentration of N protein used in each binding reaction was 3.5 × 10 −6 M. Reaction mixtures were assembled in 100 mM NaCl with 5 mM MgCl 2 in addition to standard reaction components as described in Material and Methods. Binding reactions were separated by sodium dodecyl sulfate–12% polyacrylamide gel electrophoresis, and unbound RNA was digested by adding 1 U of <t>RNase</t> V1. Signals were imaged with the Molecular Dynamics Storm PhosphorImager and quantified using ImageQuaNT version 4.2 software (Molecular Dynamics). The RNAs used to form the complexes are HTNV S-segment vRNA (lane 1), ORF RNA (lane 2), minipan RNA (lane 3), and Δ12 RNA (lane 4).
Rnase V1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rnase v1/product/GE Healthcare
Average 91 stars, based on 5 article reviews
Price from $9.99 to $1999.99
rnase v1 - by Bioz Stars, 2020-07
91/100 stars

Images

1) Product Images from "cis-Acting Signals in Encapsidation of Hantaan Virus S-Segment Viral Genomic RNA by Its N Protein"

Article Title: cis-Acting Signals in Encapsidation of Hantaan Virus S-Segment Viral Genomic RNA by Its N Protein

Journal: Journal of Virology

doi: 10.1128/JVI.75.6.2646-2652.2001

Analysis of the complex formed between HTNV N protein and deletion RNAs by UV cross-linking analysis. The concentration of N protein used in each binding reaction was 3.5 × 10 −6 M. Reaction mixtures were assembled in 100 mM NaCl with 5 mM MgCl 2 in addition to standard reaction components as described in Material and Methods. Binding reactions were separated by sodium dodecyl sulfate–12% polyacrylamide gel electrophoresis, and unbound RNA was digested by adding 1 U of RNase V1. Signals were imaged with the Molecular Dynamics Storm PhosphorImager and quantified using ImageQuaNT version 4.2 software (Molecular Dynamics). The RNAs used to form the complexes are HTNV S-segment vRNA (lane 1), ORF RNA (lane 2), minipan RNA (lane 3), and Δ12 RNA (lane 4).
Figure Legend Snippet: Analysis of the complex formed between HTNV N protein and deletion RNAs by UV cross-linking analysis. The concentration of N protein used in each binding reaction was 3.5 × 10 −6 M. Reaction mixtures were assembled in 100 mM NaCl with 5 mM MgCl 2 in addition to standard reaction components as described in Material and Methods. Binding reactions were separated by sodium dodecyl sulfate–12% polyacrylamide gel electrophoresis, and unbound RNA was digested by adding 1 U of RNase V1. Signals were imaged with the Molecular Dynamics Storm PhosphorImager and quantified using ImageQuaNT version 4.2 software (Molecular Dynamics). The RNAs used to form the complexes are HTNV S-segment vRNA (lane 1), ORF RNA (lane 2), minipan RNA (lane 3), and Δ12 RNA (lane 4).

Techniques Used: Concentration Assay, Binding Assay, Polyacrylamide Gel Electrophoresis, Software

2) Product Images from "Conserved features of Y RNAs: a comparison of experimentally derived secondary structures"

Article Title: Conserved features of Y RNAs: a comparison of experimentally derived secondary structures

Journal: Nucleic Acids Research

doi:

Summary of Y4 RNA probing data. ( A ) Summarized results of enzymatic/chemical probing on hY4 RNA, depicted on the secondary structure that was obtained by combining the experimental results with MFOLD predictions. For clarity the single-stranded (RNase T1 in blue, RNase T2 in red and RNase A in green) and double-stranded (RNase V1 in black) data are separately illustrated on the secondary structure. It should be emphasized that for large parts of the molecule, e.g. L1 and S3, alternative secondary structures are also feasible. Dotted lines represent weak cleavages, moderate cleavages are indicated by open arrowheads, while strong cleavages are indicated by filled arrowheads (see key). The DMS probing results are indicated by circles (dotted line, weak reactivity; bold line, moderate reactivity; filled circle, strong reactivity). Asterisks indicate stops of reverse transcriptase occurring in the control incubations. The oligonucleotide used for reverse transcription is complementary to the region 76–94. ( B ) Summarized probing data for xY4 (left panel) and iY4 (right panel) depicted on the respective secondary structures that were obtained by combining the experimental results with MFOLD predictions. RNase T2 cleavages are indicated in red and RNase V1 cleavages are indicated in black. Dotted lines represent weak cleavages; moderate cleavages are indicated with open arrowheads; and strong cleavages are indicated by filled arrowheads (see key).
Figure Legend Snippet: Summary of Y4 RNA probing data. ( A ) Summarized results of enzymatic/chemical probing on hY4 RNA, depicted on the secondary structure that was obtained by combining the experimental results with MFOLD predictions. For clarity the single-stranded (RNase T1 in blue, RNase T2 in red and RNase A in green) and double-stranded (RNase V1 in black) data are separately illustrated on the secondary structure. It should be emphasized that for large parts of the molecule, e.g. L1 and S3, alternative secondary structures are also feasible. Dotted lines represent weak cleavages, moderate cleavages are indicated by open arrowheads, while strong cleavages are indicated by filled arrowheads (see key). The DMS probing results are indicated by circles (dotted line, weak reactivity; bold line, moderate reactivity; filled circle, strong reactivity). Asterisks indicate stops of reverse transcriptase occurring in the control incubations. The oligonucleotide used for reverse transcription is complementary to the region 76–94. ( B ) Summarized probing data for xY4 (left panel) and iY4 (right panel) depicted on the respective secondary structures that were obtained by combining the experimental results with MFOLD predictions. RNase T2 cleavages are indicated in red and RNase V1 cleavages are indicated in black. Dotted lines represent weak cleavages; moderate cleavages are indicated with open arrowheads; and strong cleavages are indicated by filled arrowheads (see key).

Techniques Used:

Secondary structure probing experiments on Y3 RNA. ( A ) Probing experiment on hY3 RNA, using RNase V1 and RNase A under different concentrations of Mg 2+ . Lanes 1–5 show probing without Mg 2+ , lanes 2 and 3 using RNase A, lanes 4 and 5 using RNase V1, and lane 1 is a control incubation in which the nuclease was omitted. Similarly, lanes 6–10 show probing under 2.5 mM Mg 2+ , lanes 11–15 under 5 mM Mg 2+ , and lanes 16–20 under 10 mM Mg 2+ (indicated above the autoradiograph). Lanes 21 and 22 show a denaturing control incubation and an incubation with RNase T1, which are used to pinpoint nucleotide positions (indicated on the right). Note that the RNase V1 cleavage products lack the 3′ phosphate group and therefore migrate slightly slower through the gel. The positions corresponding to the various stem and loop structures as depicted in Figure 1A are shown on the left. Data for the overexposed regions were evaluated using shorter exposures. ( B ) Probing hY3 RNA with DMS at 0°C (lanes 1, 3 and 5) and 20°C (lanes 2, 4 and 6). Sites of modification were detected using primer extension. Parallel control incubations without DMS were used to detect stops of reverse transcriptase (lanes 1 and 2). The concentration of DMS is increased from lanes 3 and 4 to 5 and 6. Nucleotide positions are indicated on the right. Data for the overexposed regions were evaluated using shorter exposures. ( C ) Probing experiment on xY3 RNA, using RNase V1 (lane 2). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 3 and 4 show a sequence ladder obtained using RNase T1 (lane 4), under denaturing conditions. Lane 3 is a control incubation. On the right the nucleotide numbering is indicated. ( D ) Probing experiment on iY3 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 represents the corresponding control incubation. On the right the nucleotide numbering is indicated.
Figure Legend Snippet: Secondary structure probing experiments on Y3 RNA. ( A ) Probing experiment on hY3 RNA, using RNase V1 and RNase A under different concentrations of Mg 2+ . Lanes 1–5 show probing without Mg 2+ , lanes 2 and 3 using RNase A, lanes 4 and 5 using RNase V1, and lane 1 is a control incubation in which the nuclease was omitted. Similarly, lanes 6–10 show probing under 2.5 mM Mg 2+ , lanes 11–15 under 5 mM Mg 2+ , and lanes 16–20 under 10 mM Mg 2+ (indicated above the autoradiograph). Lanes 21 and 22 show a denaturing control incubation and an incubation with RNase T1, which are used to pinpoint nucleotide positions (indicated on the right). Note that the RNase V1 cleavage products lack the 3′ phosphate group and therefore migrate slightly slower through the gel. The positions corresponding to the various stem and loop structures as depicted in Figure 1A are shown on the left. Data for the overexposed regions were evaluated using shorter exposures. ( B ) Probing hY3 RNA with DMS at 0°C (lanes 1, 3 and 5) and 20°C (lanes 2, 4 and 6). Sites of modification were detected using primer extension. Parallel control incubations without DMS were used to detect stops of reverse transcriptase (lanes 1 and 2). The concentration of DMS is increased from lanes 3 and 4 to 5 and 6. Nucleotide positions are indicated on the right. Data for the overexposed regions were evaluated using shorter exposures. ( C ) Probing experiment on xY3 RNA, using RNase V1 (lane 2). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 3 and 4 show a sequence ladder obtained using RNase T1 (lane 4), under denaturing conditions. Lane 3 is a control incubation. On the right the nucleotide numbering is indicated. ( D ) Probing experiment on iY3 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 represents the corresponding control incubation. On the right the nucleotide numbering is indicated.

Techniques Used: Incubation, Autoradiography, Modification, Concentration Assay, Sequencing

Summary of Y3 RNA probing data. ( A ) Summarized results of enzymatic/chemical probing of hY3 RNA, depicted on two alternative secondary structures that were obtained by combining the experimental results with MFOLD predictions. For clarity the single-stranded (RNase T1 in blue, RNase T2 in red and RNase A in green) and double-stranded (RNase V1 in black) data are separately illustrated on the secondary structure drawn on the right, while the DMS data are omitted from the structure on the left. It should be emphasized that for the ‘upper’ part of the molecule additional alternative secondary structures are feasible. The experimental data suggest that at least some of these structures occur simultaneously. Dotted lines represent weak cleavages; open arrowheads indicate moderate cleavages; filled arrowheads indicate strong cleavages (see key). The DMS probing results (20°C) are indicated by circles (dotted line, weak reactivity; bold line, moderate reactivity; filled circle, strong reactivity). Asterisks indicate stops of reverse transcriptase occurring in the control incubations. The oligonucleotide used for reverse transcription is complementary to the region 82–101. ( B ) Summarized probing data for xY3 (left panel) and iY3 (right panel) depicted on the respective secondary structures that were obtained by combining the experimental results with MFOLD predictions. RNase T2 cleavages are indicated in red and RNase V1 cleavages are indicated in black. Dotted lines represent weak cleavages; moderate cleavages are indicated with open arrowheads; and strong cleavages are indicated by filled arrowheads (see key).
Figure Legend Snippet: Summary of Y3 RNA probing data. ( A ) Summarized results of enzymatic/chemical probing of hY3 RNA, depicted on two alternative secondary structures that were obtained by combining the experimental results with MFOLD predictions. For clarity the single-stranded (RNase T1 in blue, RNase T2 in red and RNase A in green) and double-stranded (RNase V1 in black) data are separately illustrated on the secondary structure drawn on the right, while the DMS data are omitted from the structure on the left. It should be emphasized that for the ‘upper’ part of the molecule additional alternative secondary structures are feasible. The experimental data suggest that at least some of these structures occur simultaneously. Dotted lines represent weak cleavages; open arrowheads indicate moderate cleavages; filled arrowheads indicate strong cleavages (see key). The DMS probing results (20°C) are indicated by circles (dotted line, weak reactivity; bold line, moderate reactivity; filled circle, strong reactivity). Asterisks indicate stops of reverse transcriptase occurring in the control incubations. The oligonucleotide used for reverse transcription is complementary to the region 82–101. ( B ) Summarized probing data for xY3 (left panel) and iY3 (right panel) depicted on the respective secondary structures that were obtained by combining the experimental results with MFOLD predictions. RNase T2 cleavages are indicated in red and RNase V1 cleavages are indicated in black. Dotted lines represent weak cleavages; moderate cleavages are indicated with open arrowheads; and strong cleavages are indicated by filled arrowheads (see key).

Techniques Used:

Secondary structure probing experiments on Y4 RNA. ( A ) Probing experiment on hY4 RNA, using RNase V1 (lanes 2–4, increasing amounts) and RNase A (lanes 5–7, increasing amounts). Lane 1 is a control incubation where RNases were omitted. Lane 8 is a control incubation and lane 9 is an RNase A incubation both under denaturing conditions to generate a sequence ladder. Nucleotide positions are indicated on the right. Note that the RNase V1 cleavage products lack the 3′ phosphate group and therefore migrate slightly slower through the gel. ( B ) Probing hY4 RNA with DMS at 0°C (lanes 1, 3 and 5) and 20°C (lanes 2, 4 and 6). Sites of modification were detected using primer extension. Parallel control incubations without DMS were used to detect stops of reverse transcriptase (lanes 1 and 2). The concentration of DMS is increased from lanes 3 and 4 to 5 and 6. Nucleotide positions are indicated on the right. ( C ) Probing experiment on xY4 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 is a control incubation. On the right the nucleotide numbering is indicated. ( D ) Probing experiment on iY4 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 is a control incubation. On the right the nucleotide numbering is indicated.
Figure Legend Snippet: Secondary structure probing experiments on Y4 RNA. ( A ) Probing experiment on hY4 RNA, using RNase V1 (lanes 2–4, increasing amounts) and RNase A (lanes 5–7, increasing amounts). Lane 1 is a control incubation where RNases were omitted. Lane 8 is a control incubation and lane 9 is an RNase A incubation both under denaturing conditions to generate a sequence ladder. Nucleotide positions are indicated on the right. Note that the RNase V1 cleavage products lack the 3′ phosphate group and therefore migrate slightly slower through the gel. ( B ) Probing hY4 RNA with DMS at 0°C (lanes 1, 3 and 5) and 20°C (lanes 2, 4 and 6). Sites of modification were detected using primer extension. Parallel control incubations without DMS were used to detect stops of reverse transcriptase (lanes 1 and 2). The concentration of DMS is increased from lanes 3 and 4 to 5 and 6. Nucleotide positions are indicated on the right. ( C ) Probing experiment on xY4 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 is a control incubation. On the right the nucleotide numbering is indicated. ( D ) Probing experiment on iY4 RNA, using RNase V1 (lane 2) and RNase T2 (lane 3). Lane 1 is the control incubation in which the enzyme was omitted. Lanes 4–6 show a sequence ladder obtained using RNase T1 (lane 5) and RNase A (lane 6), under denaturing conditions. Lane 4 is a control incubation. On the right the nucleotide numbering is indicated.

Techniques Used: Incubation, Sequencing, Modification, Concentration Assay

3) Product Images from "Putative intermediary stages for the molecular evolution from a ribozyme to a catalytic RNP"

Article Title: Putative intermediary stages for the molecular evolution from a ribozyme to a catalytic RNP

Journal: Nucleic Acids Research

doi:

( A ) RNA footprints with RNase V1 in the presence of pepA or its mutants. Autoradiograms are shown for RNase V1 footprints of the boxB (upper) and RRE (lower) regions of boxB-11nt/RRE. Dots indicate sites that show protection from RNase V1. ( B ) Degree of RNase V1 cleavage on the boxB or RRE region of boxB-11nr/RRE in the presence of pepA or its mutants. ( C ) Degree of RNase V1 cleavage on the boxB or RRE region of boxB-11nr/RRE and its mutants in the presence or absence of pepA.
Figure Legend Snippet: ( A ) RNA footprints with RNase V1 in the presence of pepA or its mutants. Autoradiograms are shown for RNase V1 footprints of the boxB (upper) and RRE (lower) regions of boxB-11nt/RRE. Dots indicate sites that show protection from RNase V1. ( B ) Degree of RNase V1 cleavage on the boxB or RRE region of boxB-11nr/RRE in the presence of pepA or its mutants. ( C ) Degree of RNase V1 cleavage on the boxB or RRE region of boxB-11nr/RRE and its mutants in the presence or absence of pepA.

Techniques Used:

4) Product Images from "Eukaryotic Initiation Factors 4G and 4A Mediate Conformational Changes Downstream of the Initiation Codon of the Encephalomyocarditis Virus Internal Ribosomal Entry Site"

Article Title: Eukaryotic Initiation Factors 4G and 4A Mediate Conformational Changes Downstream of the Initiation Codon of the Encephalomyocarditis Virus Internal Ribosomal Entry Site

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.23.2.687-698.2003

Chemical and enzymatic footprinting analyses of ribonucleoprotein complexes assembled on the EMCV IRES. (A) Chemical foot-printing of the wild-type and ΔJ5 and ΔK2′ mutant forms of EMCV (nt 315 to 1155) RNA in binary complexes formed with eIF4GII 745-1003 . (B) Chemical and enzymatic footprinting of wild-type EMCV (nt 315 to 1155) RNA in complexes formed with eIF4GI 737-1600 , eIF4A, and eIF4B, as indicated. Polyacrylamide-urea gel fractionation of cDNA products after primer extension by AMV-RT shows the sensitivity of the IRES to either DMS or RNase V1, either alone or complexed with initiation factors, as indicated. cDNA products obtained after primer extension of untreated EMCV RNAs are shown in lanes 1, 4, and 7 of panel A and in lane 1 of panel B. The positions of protected residues are indicated by the symbols shown in the key at bottom left; some sites of protection on mutant mRNAs are indicated by arrowheads on panel A. A dideoxynucleotide sequence generated with the same primer was run in parallel (lanes C, T, A, and G in panel A), and the positions of EMCV nucleotides at 50-nt intervals are indicated to the left of both panels. The full-length cDNA extension product is marked E.
Figure Legend Snippet: Chemical and enzymatic footprinting analyses of ribonucleoprotein complexes assembled on the EMCV IRES. (A) Chemical foot-printing of the wild-type and ΔJ5 and ΔK2′ mutant forms of EMCV (nt 315 to 1155) RNA in binary complexes formed with eIF4GII 745-1003 . (B) Chemical and enzymatic footprinting of wild-type EMCV (nt 315 to 1155) RNA in complexes formed with eIF4GI 737-1600 , eIF4A, and eIF4B, as indicated. Polyacrylamide-urea gel fractionation of cDNA products after primer extension by AMV-RT shows the sensitivity of the IRES to either DMS or RNase V1, either alone or complexed with initiation factors, as indicated. cDNA products obtained after primer extension of untreated EMCV RNAs are shown in lanes 1, 4, and 7 of panel A and in lane 1 of panel B. The positions of protected residues are indicated by the symbols shown in the key at bottom left; some sites of protection on mutant mRNAs are indicated by arrowheads on panel A. A dideoxynucleotide sequence generated with the same primer was run in parallel (lanes C, T, A, and G in panel A), and the positions of EMCV nucleotides at 50-nt intervals are indicated to the left of both panels. The full-length cDNA extension product is marked E.

Techniques Used: Footprinting, Mutagenesis, Fractionation, Sequencing, Generated

Related Articles

Incubation:

Article Title: Putative intermediary stages for the molecular evolution from a ribozyme to a catalytic RNP
Article Snippet: .. After preincubation for 5 min, RNA–protein complexes were digested by adding 2 µl of RNase V1 (0.4 U/ml; Amersham Pharmacia Biotech), followed by incubation for 10 min. .. The RNAs were extracted with phenol and recovered by ethanol precipitation.

Article Title: Design and development of a catalytic ribonucleoprotein
Article Snippet: .. After pre-incubation for 5 min, M12 RNA or RNA–protein complexes were digested by addition of 1 µl of RNase V1 (0.5 U/ml; Amersham Pharmacia Biotech) or incubated with 1 µl of DMS diluted to 1:600, followed by incubation for 10 min at 37°C. ..

other:

Article Title: Secondary structure of two regions in expansion segments ES3 and ES6 with the potential of forming a tertiary interaction in eukaryotic 40S ribosomal subunits
Article Snippet: Ribonuclease V1 and cDNA oligonucleotides were from Amersham Pharmacia Biotech.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    GE Healthcare rnase cocktail
    Rnase Cocktail, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase cocktail/product/GE Healthcare
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rnase cocktail - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    Image Search Results