Structured Review

Thermo Fisher rnase a
RNase and DNase analysis of major satellite species. Total RNA from NIH/3T3 cells was treated with indicated enzymes for 30 min at 37°, phenol:chloroform extracted, ethanol precipitated and glyoxylated prior to electrophoresis, then blotted and probed for major satellite sequences, ethidium staining of ribosomal RNA is shown as a control. A: Samples treated in water without and with RNase A. B: Samples treated in NEBuffer 3 with indicated ribonucleases. C: Samples treated in RQ1 DNase buffer without and with DNase I. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, p values calculated using a one-way ANOVA (n = 4) for B and Student’s  t -test (n = 3) for C.
Rnase A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rnase a/product/Thermo Fisher
Average 90 stars, based on 135 article reviews
Price from $9.99 to $1999.99
rnase a - by Bioz Stars, 2020-01
90/100 stars

Images

1) Product Images from "Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A"

Article Title: Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A

Journal: PLoS ONE

doi: 10.1371/journal.pone.0115008

RNase and DNase analysis of major satellite species. Total RNA from NIH/3T3 cells was treated with indicated enzymes for 30 min at 37°, phenol:chloroform extracted, ethanol precipitated and glyoxylated prior to electrophoresis, then blotted and probed for major satellite sequences, ethidium staining of ribosomal RNA is shown as a control. A: Samples treated in water without and with RNase A. B: Samples treated in NEBuffer 3 with indicated ribonucleases. C: Samples treated in RQ1 DNase buffer without and with DNase I. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, p values calculated using a one-way ANOVA (n = 4) for B and Student’s  t -test (n = 3) for C.
Figure Legend Snippet: RNase and DNase analysis of major satellite species. Total RNA from NIH/3T3 cells was treated with indicated enzymes for 30 min at 37°, phenol:chloroform extracted, ethanol precipitated and glyoxylated prior to electrophoresis, then blotted and probed for major satellite sequences, ethidium staining of ribosomal RNA is shown as a control. A: Samples treated in water without and with RNase A. B: Samples treated in NEBuffer 3 with indicated ribonucleases. C: Samples treated in RQ1 DNase buffer without and with DNase I. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, p values calculated using a one-way ANOVA (n = 4) for B and Student’s t -test (n = 3) for C.

Techniques Used: Electrophoresis, Staining

Inhibition of DNA removal by RNase A. A: 0.5 ng major satellite PCR product was mixed with 1 µg NIH/3T3 RNA (lanes 1, 2) or with 1 µg DNA molecular weight marker (lanes 3, 4), and treated without or with RNase A followed by phenol:chloroform extraction and analysis as in Fig. 1. B: Identical to A, except that samples were incubated with 20 µg proteinase K for 15 min at 37° after RNase A treatment and phenol:chloroform extraction was omitted. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, analysis by one-way ANOVA (n = 3), differences in B are not significant.
Figure Legend Snippet: Inhibition of DNA removal by RNase A. A: 0.5 ng major satellite PCR product was mixed with 1 µg NIH/3T3 RNA (lanes 1, 2) or with 1 µg DNA molecular weight marker (lanes 3, 4), and treated without or with RNase A followed by phenol:chloroform extraction and analysis as in Fig. 1. B: Identical to A, except that samples were incubated with 20 µg proteinase K for 15 min at 37° after RNase A treatment and phenol:chloroform extraction was omitted. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, analysis by one-way ANOVA (n = 3), differences in B are not significant.

Techniques Used: Inhibition, Polymerase Chain Reaction, Molecular Weight, Marker, Incubation

RNase A activity on DNA. A, B: 0.5 ng major satellite PCR product was mixed with 1 µg NIH/3T3 total RNA and analysed as in Fig. 1. A: Treatment with RNase A from different manufacturers. B: Treatment with RNase A in different buffers. C: 50 ng DNA molecular weight marker was mixed with 1 µg NIH/3T3 total RNA and treated without or with RNase A, purified as in Fig. 1 and separated on a non-denaturing 1xTBE gel before blotting and probing for the molecular weight marker. D: 32 P-labelled 50 bp ladder mixed with 1 µg RNA was treated without or with RNase A for 30 min at 37°, directly separated on an 8% PAGE gel and exposed to a phosphorimaging screen. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, p values were calculated by one-way ANOVA (A, B) or Student’s t -test (C, D), n = 3 in all cases.
Figure Legend Snippet: RNase A activity on DNA. A, B: 0.5 ng major satellite PCR product was mixed with 1 µg NIH/3T3 total RNA and analysed as in Fig. 1. A: Treatment with RNase A from different manufacturers. B: Treatment with RNase A in different buffers. C: 50 ng DNA molecular weight marker was mixed with 1 µg NIH/3T3 total RNA and treated without or with RNase A, purified as in Fig. 1 and separated on a non-denaturing 1xTBE gel before blotting and probing for the molecular weight marker. D: 32 P-labelled 50 bp ladder mixed with 1 µg RNA was treated without or with RNase A for 30 min at 37°, directly separated on an 8% PAGE gel and exposed to a phosphorimaging screen. For quantification, error bars represent ±1 s.d., y-axes in arbitrary units, p values were calculated by one-way ANOVA (A, B) or Student’s t -test (C, D), n = 3 in all cases.

Techniques Used: Activity Assay, Polymerase Chain Reaction, Molecular Weight, Marker, Purification, Polyacrylamide Gel Electrophoresis

Re-partitioning of RNase-treated DNA. A, B: 5 ng 32 P-labelled major satellite PCR product mixed with 1 µg of NIH/3T3 RNA was incubated without or with RNase A for 30 min at 37°. Reactions were then diluted to 100 µl with water and extracted with 100 µl phenol:chloroform. A: 10 µl of the aqueous phase was analysed by electrophoresis in a non-denaturing 1xTBE gel which was dried and exposed to a phosphorimaging screen. B: Radioactivity present in the aqueous and phenol phases quantitated using a Geiger counter. C: DNA loss after RNase A treatment with phenol:chloroform extraction at pH 7 or pH 8. D: Activity of RNase A in phenol:chloroform. 1 µl 1 mg/ml RNase A was added to 100 µl phenol:chloroform and vortexed. 1 µl 1 mg/ml NIH/3T3 RNA was added, vortexed and reactions incubated at 37° for 30 min, followed by extraction with 100 µl water, precipitation and gel electrophoresis. For quantification, error bars represent ±1 s.d, y-axes in arbitrary units for A, D or Bequerels in B, analysis by Student’s t -test, n = 3.
Figure Legend Snippet: Re-partitioning of RNase-treated DNA. A, B: 5 ng 32 P-labelled major satellite PCR product mixed with 1 µg of NIH/3T3 RNA was incubated without or with RNase A for 30 min at 37°. Reactions were then diluted to 100 µl with water and extracted with 100 µl phenol:chloroform. A: 10 µl of the aqueous phase was analysed by electrophoresis in a non-denaturing 1xTBE gel which was dried and exposed to a phosphorimaging screen. B: Radioactivity present in the aqueous and phenol phases quantitated using a Geiger counter. C: DNA loss after RNase A treatment with phenol:chloroform extraction at pH 7 or pH 8. D: Activity of RNase A in phenol:chloroform. 1 µl 1 mg/ml RNase A was added to 100 µl phenol:chloroform and vortexed. 1 µl 1 mg/ml NIH/3T3 RNA was added, vortexed and reactions incubated at 37° for 30 min, followed by extraction with 100 µl water, precipitation and gel electrophoresis. For quantification, error bars represent ±1 s.d, y-axes in arbitrary units for A, D or Bequerels in B, analysis by Student’s t -test, n = 3.

Techniques Used: Polymerase Chain Reaction, Incubation, Electrophoresis, Radioactivity, Activity Assay, Nucleic Acid Electrophoresis

2) Product Images from "Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile"

Article Title: Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkt1009

Analysis of tRNA 1 Ile purified from Bacillus subtilis wild-type and mutant strain JJS80. ( A ) Characterization of purified tRNA 1 from B. subtilis wild-type (WT; lanes 1–4) and tRNA 1 from JJS80 (lanes 5–8) by RNase A and T1 analysis. The 5′- 32 P-labeled tRNA was partially digested with 0.004 U of RNase A (lanes 2 and 7) and 10 U of RNase T1 (lanes 3 and 6). Lanes 4 and 5 show a partial alkali digest; lanes 1 and 8 show undigested control reactions. 32 P-labeled fragments were separated by denaturing PAGE and visualized by autoradiography. ( B ) Analysis of purified tRNA 1 and tRNA 1 by 15% native PAGE. tRNAs were visualized by staining with ethidium bromide (EtBr) and northern hybridization using a universal tRNA 1 Ile probe. 0.01 A 260 of total tRNA were applied per lane.
Figure Legend Snippet: Analysis of tRNA 1 Ile purified from Bacillus subtilis wild-type and mutant strain JJS80. ( A ) Characterization of purified tRNA 1 from B. subtilis wild-type (WT; lanes 1–4) and tRNA 1 from JJS80 (lanes 5–8) by RNase A and T1 analysis. The 5′- 32 P-labeled tRNA was partially digested with 0.004 U of RNase A (lanes 2 and 7) and 10 U of RNase T1 (lanes 3 and 6). Lanes 4 and 5 show a partial alkali digest; lanes 1 and 8 show undigested control reactions. 32 P-labeled fragments were separated by denaturing PAGE and visualized by autoradiography. ( B ) Analysis of purified tRNA 1 and tRNA 1 by 15% native PAGE. tRNAs were visualized by staining with ethidium bromide (EtBr) and northern hybridization using a universal tRNA 1 Ile probe. 0.01 A 260 of total tRNA were applied per lane.

Techniques Used: Purification, Mutagenesis, Labeling, Polyacrylamide Gel Electrophoresis, Autoradiography, Clear Native PAGE, Staining, Northern Blot, Hybridization

3) Product Images from "Vesicular stomatitis virus inhibits mitotic progression and triggers cell death"

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death

Journal: EMBO Reports

doi: 10.1038/embor.2009.179

VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.
Figure Legend Snippet: VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.

Techniques Used: Incubation, Recombinant, SDS Page, Immunoprecipitation, Immunofluorescence, Microscopy, Polyacrylamide Gel Electrophoresis

4) Product Images from "FMRP recruitment of β‐catenin to the translation pre‐initiation complex represses translation"

Article Title: FMRP recruitment of β‐catenin to the translation pre‐initiation complex represses translation

Journal: EMBO Reports

doi: 10.15252/embr.201745536

β‐Catenin and FMRP are co‐localized in the pre‐initiation complex Co‐sedimentation of protein/RNA complexes on a 10‐50% sucrose gradient. Cell lysates generated from the cells transfected with either siRNA targeting fmr1 or its scrambled control were ultracentrifuged in sucrose gradients; peaks corresponding to the 40S and 60S subunits, 80S monosome, and polysomes were detected by UV absorbance at 254 nm, and indicated proteins in these fractions were detected by Western blot. Corresponding total cell lysate was used as input. (i) eIF4E preferentially binds to the 5′cap (m 7 GTP) of mRNAs and recruits the pre‐initiation complex to that site. (ii) m 7 GTP‐agarose beads. (i) Precipitated proteins in A10 cell lysates with m 7 GTP‐agarose beads were identified by Western blot analysis. eIF4E and tubulin were shown as positive and negative controls, respectively. Total lysates were used as input. (ii) A10 cell lysates were incubated with GTP‐agarose beads, and none of the proteins tested were precipitated with the beads. Total lysates were used as input control. (i) HEK 293T cells were transfected with empty vector or Flag‐FMRP, and lysates were subjected to m 7 GTP‐agarose pull‐down as in (C). (ii) HEK 293T cells were transfected with empty vector or Flag‐FMRP, and lysates were subjected to GTP‐agarose pull‐down as in (i). HEK 293T cells were transfected with either siRNAs targeting fmr1 or scrambled control and were subjected to m 7 GTP‐agarose pull‐downs as in (C). HEK 293T cells lysates were subjected to m 7 GTP‐agarose pull‐downs as in (C) in the presence or absence of RNase A (10 μg/ml). RNA was extracted from parallel lysates, and RNA content was analyzed by agarose gel electrophoresis.
Figure Legend Snippet: β‐Catenin and FMRP are co‐localized in the pre‐initiation complex Co‐sedimentation of protein/RNA complexes on a 10‐50% sucrose gradient. Cell lysates generated from the cells transfected with either siRNA targeting fmr1 or its scrambled control were ultracentrifuged in sucrose gradients; peaks corresponding to the 40S and 60S subunits, 80S monosome, and polysomes were detected by UV absorbance at 254 nm, and indicated proteins in these fractions were detected by Western blot. Corresponding total cell lysate was used as input. (i) eIF4E preferentially binds to the 5′cap (m 7 GTP) of mRNAs and recruits the pre‐initiation complex to that site. (ii) m 7 GTP‐agarose beads. (i) Precipitated proteins in A10 cell lysates with m 7 GTP‐agarose beads were identified by Western blot analysis. eIF4E and tubulin were shown as positive and negative controls, respectively. Total lysates were used as input. (ii) A10 cell lysates were incubated with GTP‐agarose beads, and none of the proteins tested were precipitated with the beads. Total lysates were used as input control. (i) HEK 293T cells were transfected with empty vector or Flag‐FMRP, and lysates were subjected to m 7 GTP‐agarose pull‐down as in (C). (ii) HEK 293T cells were transfected with empty vector or Flag‐FMRP, and lysates were subjected to GTP‐agarose pull‐down as in (i). HEK 293T cells were transfected with either siRNAs targeting fmr1 or scrambled control and were subjected to m 7 GTP‐agarose pull‐downs as in (C). HEK 293T cells lysates were subjected to m 7 GTP‐agarose pull‐downs as in (C) in the presence or absence of RNase A (10 μg/ml). RNA was extracted from parallel lysates, and RNA content was analyzed by agarose gel electrophoresis.

Techniques Used: Sedimentation, Generated, Transfection, Western Blot, Incubation, Plasmid Preparation, Agarose Gel Electrophoresis

β‐Catenin interaction with FMRP (Left panel) A10 cells were transfected with Flag‐FMRP for 3 h then cultured for 24 h prior to fixation and staining for β‐catenin (green), Flag (FMRP, red), and nucleus (blue). (Bottom right panel) A line was drawn through a representative cell to indicate relative intensity of RGB signals. (Top right panel) An orthogonal projection was generated by a Z ‐stack (240 nm interval) image set using the ZEN program (Zeiss) depicting the region surrounding the nucleus in the XY, XZ, and YZ planes. (i) HEK 293T cells were transfected with Flag‐FMRP as in (A), and endogenous β‐catenin protein complexes were immunoprecipitated with β‐catenin antibody. Co‐immunoprecipitated Flag‐FMRP protein was detected by Western blot analysis with the Flag antibody. Non‐programmed rabbit IgG was used as a control. (ii) Primary VSMCs were harvested and subjected to an endogenous coIP analysis with either β‐catenin or FMRP antibody. The corresponding protein was detected in the precipitated immunocomplex by Western blot. Non‐programmed rabbit IgG was used as a control. (i) Purified 6x‐His‐FMRP and various regions of β‐catenin proteins fused to GST were incubated with glutathione‐agarose beads. Purified GST or GST‐fusion proteins were visualized by Western blotting using the GST antibody, and sizes of corresponding GST‐fusion proteins were confirmed. Pulled‐down FMRP proteins by GST fused β‐catenin were detected by immunoblotting with the FMRP antibody. (ii) Interaction between FMRP and β‐catenin was assessed in the presence or absence of RNA. HEK 293T cells were subjected to an endogenous coIP analysis using either β‐catenin or FMRP antibody, in the presence or absence of RNase A (10 μg/ml). The corresponding protein was detected in the precipitated immunocomplex by Western blot, and non‐programmed rabbit IgG was used as a control. RNA was extracted from parallel cultured cells, and total RNA content was analyzed by agarose gel electrophoresis in the presence or absence of RNase A. (Top panel) A10 cells were transfected with GBP‐Lifeact and either EYFP‐β‐catenin, Flag‐FMRP, or mCherry‐β‐catenin. The cells were fixed and subjected to either fluorescence microscopy or immunofluorescent staining for Flag (red), as indicated. (Bottom panel) A10 cells were transfected with GBP‐Lifeact similar to above, but with both EYFP‐β‐catenin and Flag‐FMRP, and the cells were subjected to fluorescent microscopy and immunofluorescent staining for Flag (red). Scale bars, 20 μm. A10 cells were transfected as in (D) with GBP‐lamin B1. Scale bars, 20 μm.
Figure Legend Snippet: β‐Catenin interaction with FMRP (Left panel) A10 cells were transfected with Flag‐FMRP for 3 h then cultured for 24 h prior to fixation and staining for β‐catenin (green), Flag (FMRP, red), and nucleus (blue). (Bottom right panel) A line was drawn through a representative cell to indicate relative intensity of RGB signals. (Top right panel) An orthogonal projection was generated by a Z ‐stack (240 nm interval) image set using the ZEN program (Zeiss) depicting the region surrounding the nucleus in the XY, XZ, and YZ planes. (i) HEK 293T cells were transfected with Flag‐FMRP as in (A), and endogenous β‐catenin protein complexes were immunoprecipitated with β‐catenin antibody. Co‐immunoprecipitated Flag‐FMRP protein was detected by Western blot analysis with the Flag antibody. Non‐programmed rabbit IgG was used as a control. (ii) Primary VSMCs were harvested and subjected to an endogenous coIP analysis with either β‐catenin or FMRP antibody. The corresponding protein was detected in the precipitated immunocomplex by Western blot. Non‐programmed rabbit IgG was used as a control. (i) Purified 6x‐His‐FMRP and various regions of β‐catenin proteins fused to GST were incubated with glutathione‐agarose beads. Purified GST or GST‐fusion proteins were visualized by Western blotting using the GST antibody, and sizes of corresponding GST‐fusion proteins were confirmed. Pulled‐down FMRP proteins by GST fused β‐catenin were detected by immunoblotting with the FMRP antibody. (ii) Interaction between FMRP and β‐catenin was assessed in the presence or absence of RNA. HEK 293T cells were subjected to an endogenous coIP analysis using either β‐catenin or FMRP antibody, in the presence or absence of RNase A (10 μg/ml). The corresponding protein was detected in the precipitated immunocomplex by Western blot, and non‐programmed rabbit IgG was used as a control. RNA was extracted from parallel cultured cells, and total RNA content was analyzed by agarose gel electrophoresis in the presence or absence of RNase A. (Top panel) A10 cells were transfected with GBP‐Lifeact and either EYFP‐β‐catenin, Flag‐FMRP, or mCherry‐β‐catenin. The cells were fixed and subjected to either fluorescence microscopy or immunofluorescent staining for Flag (red), as indicated. (Bottom panel) A10 cells were transfected with GBP‐Lifeact similar to above, but with both EYFP‐β‐catenin and Flag‐FMRP, and the cells were subjected to fluorescent microscopy and immunofluorescent staining for Flag (red). Scale bars, 20 μm. A10 cells were transfected as in (D) with GBP‐lamin B1. Scale bars, 20 μm.

Techniques Used: Transfection, Cell Culture, Staining, Generated, Immunoprecipitation, Western Blot, Co-Immunoprecipitation Assay, Purification, Incubation, Agarose Gel Electrophoresis, Fluorescence, Microscopy

5) Product Images from "RNA-binding protein FXR1 is presented in rat brain in amyloid form"

Article Title: RNA-binding protein FXR1 is presented in rat brain in amyloid form

Journal: Scientific Reports

doi: 10.1038/s41598-019-55528-6

FXR1 colocalizes with amyloid-specific dye Thioflavin S in cortical neurons and colocalizes with mRNA molecules resistant to RNAse treatment. ( a ) FXR1 is present in the perinuclear cytoplasm of cortical neurons and colocalizes with Thioflavin S. See also Supplementary Fig.   S6 . ( b ) FXR1 colocalizes with some portion of mRNA in the cytoplasm of cortical neurons. ( c ) mRNAs that are colocalized with FXR1 are detected after treatment with RNAse A, whereas other mRNAs degrade. Immunohistochemistry and fluorescent  in situ  hybridization on the cryosections of the rat brain cortex were carried out as described in “Materials and methods”. Scale bar for sections ( a – c ) is 20 µm.
Figure Legend Snippet: FXR1 colocalizes with amyloid-specific dye Thioflavin S in cortical neurons and colocalizes with mRNA molecules resistant to RNAse treatment. ( a ) FXR1 is present in the perinuclear cytoplasm of cortical neurons and colocalizes with Thioflavin S. See also Supplementary Fig.  S6 . ( b ) FXR1 colocalizes with some portion of mRNA in the cytoplasm of cortical neurons. ( c ) mRNAs that are colocalized with FXR1 are detected after treatment with RNAse A, whereas other mRNAs degrade. Immunohistochemistry and fluorescent in situ hybridization on the cryosections of the rat brain cortex were carried out as described in “Materials and methods”. Scale bar for sections ( a – c ) is 20 µm.

Techniques Used: Immunohistochemistry, In Situ Hybridization

6) Product Images from "Protein-imprinted polysiloxane scaffolds"

Article Title: Protein-imprinted polysiloxane scaffolds

Journal:

doi: 10.1016/j.actbio.2007.01.003

Competitive binding of protein to scaffolds imprinted with (a) 0.05 mg and (b) 0.1 mg of RNase A.
Figure Legend Snippet: Competitive binding of protein to scaffolds imprinted with (a) 0.05 mg and (b) 0.1 mg of RNase A.

Techniques Used: Binding Assay

Amount of RNase A released from the surface of scaffolds after 3 and 24 h of digestion. The inset is an expanded view of low loading region.
Figure Legend Snippet: Amount of RNase A released from the surface of scaffolds after 3 and 24 h of digestion. The inset is an expanded view of low loading region.

Techniques Used:

7) Product Images from "Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity"

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2019.02753

The growth-promoting effect of eDNA on M. bovis . Comparative growth of M. bovis under axenic and cell culture conditions (A) . RM16 proliferation was monitored in SP4 medium (SP4), cell culture medium (DMEM), and cell culture medium supplemented with 10 μg/ml calf thymus DNA (DMEMD). Mycoplasma titers (log CFU/ml) were determined by CFU titrations. The cytopathic effect induced by M. bovis upon co-incubation with host cells (B) . EBL cells (10 4 cells) were inoculated with RM16 at an MOI of 2 (RM16) or mock-infected (Mock). After 72 h of co-incubation, monolayers were stained with crystal violet and survival cells were estimated by measuring the optical density at 590 nm (OD 590). When indicated, DMEM-based medium was supplemented with 10 μg/ml calf thymus DNA (eDNA). Calf thymus DNA was subjected to the following enzymatic treatments: RNase A (RNase), Proteinase K (ProtK) and DNase I (DNase) digestion (see section “Materials and Methods”). The asterisk indicates that polynucleotides were removed from DNase I digestion products (DNase*). Infected and mock-infected samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing OD590 values of RM16-infected samples to those of mock-infected samples ( *** p
Figure Legend Snippet: The growth-promoting effect of eDNA on M. bovis . Comparative growth of M. bovis under axenic and cell culture conditions (A) . RM16 proliferation was monitored in SP4 medium (SP4), cell culture medium (DMEM), and cell culture medium supplemented with 10 μg/ml calf thymus DNA (DMEMD). Mycoplasma titers (log CFU/ml) were determined by CFU titrations. The cytopathic effect induced by M. bovis upon co-incubation with host cells (B) . EBL cells (10 4 cells) were inoculated with RM16 at an MOI of 2 (RM16) or mock-infected (Mock). After 72 h of co-incubation, monolayers were stained with crystal violet and survival cells were estimated by measuring the optical density at 590 nm (OD 590). When indicated, DMEM-based medium was supplemented with 10 μg/ml calf thymus DNA (eDNA). Calf thymus DNA was subjected to the following enzymatic treatments: RNase A (RNase), Proteinase K (ProtK) and DNase I (DNase) digestion (see section “Materials and Methods”). The asterisk indicates that polynucleotides were removed from DNase I digestion products (DNase*). Infected and mock-infected samples were treated identically. Data are the means of at least three independent assays. Standard deviations are indicated by error bars. p -values were determined by using two-sided independent sample t tests and comparing OD590 values of RM16-infected samples to those of mock-infected samples ( *** p

Techniques Used: Cell Culture, Incubation, Infection, Staining

8) Product Images from "CRISPR RNA-guided DNA cleavage by reconstituted Type I-A immune effector complexes"

Article Title: CRISPR RNA-guided DNA cleavage by reconstituted Type I-A immune effector complexes

Journal: Extremophiles : life under extreme conditions

doi: 10.1007/s00792-018-1057-0

Identification of direct interactions between crRNA and Csa proteins by UV crosslinking. SDS-polyacryladmide gel showing Coomassie blue (G-250) stained Csa proteins and RNase T1 and/or RNase A in each reaction and corresponding autoradiograph images (rad)
Figure Legend Snippet: Identification of direct interactions between crRNA and Csa proteins by UV crosslinking. SDS-polyacryladmide gel showing Coomassie blue (G-250) stained Csa proteins and RNase T1 and/or RNase A in each reaction and corresponding autoradiograph images (rad)

Techniques Used: Staining, Autoradiography

9) Product Images from "Intracellular MicroRNA Quantification in Intact Cells: A Novel Strategy based on Reduced Graphene Oxide Based Fluorescence Quenching"

Article Title: Intracellular MicroRNA Quantification in Intact Cells: A Novel Strategy based on Reduced Graphene Oxide Based Fluorescence Quenching

Journal: MRS communications

doi: 10.1557/mrc.2018.120

Sensitivity of fluorescence enhancements and the RNase A cleavage efficiency associated fluorescence enhancements of Cy5-antimiR-21 and Cy5-antimiR-21-miR-21 hybrid with and without rGO measured by spectroscopy; (a) ss-Cy5-antimiR-21 measured for fluorescence
Figure Legend Snippet: Sensitivity of fluorescence enhancements and the RNase A cleavage efficiency associated fluorescence enhancements of Cy5-antimiR-21 and Cy5-antimiR-21-miR-21 hybrid with and without rGO measured by spectroscopy; (a) ss-Cy5-antimiR-21 measured for fluorescence

Techniques Used: Fluorescence, Spectroscopy

10) Product Images from "A cis-replication element functions in both orientations to enhance replication of Turnip crinkle virus"

Article Title: A cis-replication element functions in both orientations to enhance replication of Turnip crinkle virus

Journal:

doi: 10.1016/j.virol.2006.03.051

Chemical and enzymatic probing of H4(+) and H4(−). TCV plus-strand transcripts (A) or minus-strand transcripts (B) were treated with DMS for 10 or 20 minutes or with RNase T1, RNase A and RNase V1 for 5 or 10 minutes. The modified or cleaved RNAs
Figure Legend Snippet: Chemical and enzymatic probing of H4(+) and H4(−). TCV plus-strand transcripts (A) or minus-strand transcripts (B) were treated with DMS for 10 or 20 minutes or with RNase T1, RNase A and RNase V1 for 5 or 10 minutes. The modified or cleaved RNAs

Techniques Used: Modification

11) Product Images from "The role of a metastable RNA secondary structure in hepatitis delta virus genotype III RNA editing"

Article Title: The role of a metastable RNA secondary structure in hepatitis delta virus genotype III RNA editing

Journal:

doi: 10.1261/rna.89306

Secondary structure analysis of MD-III-2 Upper and Lower RNAs. ( A ) Gel purified Upper (U) and Lower (L) RNAs were either untreated (lanes −) or digested with RNase T1 or RNase A, (lanes ++, 1 unit; lanes +, 0.1 units). The gel shown is representative
Figure Legend Snippet: Secondary structure analysis of MD-III-2 Upper and Lower RNAs. ( A ) Gel purified Upper (U) and Lower (L) RNAs were either untreated (lanes −) or digested with RNase T1 or RNase A, (lanes ++, 1 unit; lanes +, 0.1 units). The gel shown is representative

Techniques Used: Purification

Related Articles

Centrifugation:

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: Upon removal of phosphate buffered saline (PBS), cells were scraped off and precipitated by centrifugation at 3000 rpm for 10 min. Pellets were resuspended in 1 ml lysis buffer (50 mM Tris–HCl pH 7.4, 100 mM NaCl, 1 mM MgCl2 , 0.1 mM CaCl2 , 1% NP-40, 0.1% SDS, 0.5% sodium deoxycholate). .. Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm.

Article Title: Differential ratio amplicons (Ramp) for the evaluation of RNA integrity extracted from complex environmental samples
Article Snippet: Enzymatic degradation by RNase I For RNAse I degradation experiment, 40 μl aliquots of DNA‐free RNA was incubated at 37°C for 40 min in the presence of increasing concentrations of RNase I (Ambion): 0 (buffer only), 2, 10, 20 and 40 Units RNase I μg−1 RNA. .. RNA was pelleted by centrifugation 16 000g for 40 min at 4°C and the pellet was washed with 480 μl ice cold 70% ethanol and pelleted by centrifugation at 16 000g for 30 min at 4°C.

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. To test the cytotoxicity of cell culture supernatants infected with M. bovis , mycoplasma cells were killed by a gentamicin treatment (400 μg/ml) for 3 h at 37°C and mycoplasma cells were depleted by 30 min centrifugation at 12,000 ×g (4°C).

Amplification:

Article Title: Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A
Article Snippet: Major satellite PCR product was amplified using Taq polymerase (NEB) and primers T7 and T3 from pCR4-Maj9-2 , digested with Eco RI and gel purified using a QIAQuick column (QIAGEN). .. RNase A (Roche 10109142001) was dissolved at 10 mg/ml in 10 mM sodium acetate pH 5.2, boiled for 15 min then neutralised by addition of Tris pH 7.4 to 0.1 M then diluted to a 1 mg/ml stock, 10 mg/ml DNase-free RNase (Thermo EN0531) was used as provided (0.1 µl per reaction).

Mass Spectrometry:

Article Title: Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
Article Snippet: For RNase A or T1 digestion analysis, 70 pmol test RNA was incubated with RNase A (20 µg, Thermo Fisher Scientific) in 10 mM Tris-HCl pH < 7, 20 µM EDTA, 10 mM NaCl for 1 h at 60°C; or RNase T1 (3400 u, Thermo Fisher Scientific) in 50 mM Tris-HCl pH 6, 1 mM EDTA for 1 h at 57°C. .. Of note, 60 pmol of digested RNA was loaded onto an integrated HPLC/ESI-MS Agilent 1200 series system equipped with a Zorbax C18 column (50 × 2.1 mm, Supelco) and eluted with a gradient of solvents A (5 mM ammonium acetate, pH 7.4) and B (5 mM ammonium acetate pH 7.4, 99% methanol) as follows: 0–15 min, 0%–15% B; 15–20 min, 15%–25% B; 20–20.5 min, 25%–100% B. High-resolution mass spectra of the HPLC-separated RNA digestion products were acquired on a Q-TOF 6250 mass spectrometer (Agilent) equipped with a Dual-ESI source.

Construct:

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: The peptidyl-tRNA construct was generated by creating a truncated mRNA lacking a stop codon directly after the Thrdx-HA sequence. .. For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Cytotoxicity Assay:

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Paragraph title: Co-cultivation of M. bovis With Bovine Lung Cells and Crystal Violet Cell Cytotoxicity Assay ... Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C.

Incubation:

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death
Article Snippet: .. For RNase A or RNAsin pre-treatments, cell lysates were pre-incubated with RNAsin (1,000 units/ml) or RNase A (50 μg/ml; Ambion, Austin, TX, USA) for 15 min at 37°C, followed by incubation on ice for 20 min. .. Supplementary information is available at EMBO reports ).

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: .. Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm. ..

Article Title: Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile
Article Snippet: .. Partial digestion of tRNA with RNase A: 1 μg of tRNA in 10 mM Tris–HCl, pH 7.5, and 1 mM EDTA was pre-incubated at 37°C for 5 min before RNase A (0.004 U; Ambion) was added and samples were incubated at 37°C for 5 min. ..

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: .. For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel. ..

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C. .. In order to determine the binding of endogenous ADARs to Dicer, Huh-7 cells were analysed by reciprocal pulldown assays using the ADAR1, ADAR2 or Dicer-specific antibody.

Article Title: Novel Structure and Unexpected RNA-Binding Ability of the C-Terminal Domain of Herpes Simplex Virus 1 Tegument Protein UL21
Article Snippet: The upper aqueous layer was mixed with 10 μg glycogen (Life Technologies), 1/10 volume 3 M sodium acetate, and 3 volumes ethanol and was incubated at −20°C for 30 min. Nucleic acids were pelleted at 16,000 × g and 4°C for 20 min and were washed with 75% ethanol before resuspension in water for analysis. .. To isolate DNA only, the complex was treated with RNase A at an RNase A (Invitrogen)/UL21C volume ratio of 1:1,000 for 20 min at 37°C prior to nucleic acid extraction with the neutral pH phenol-chloroform mixture.

Article Title: Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
Article Snippet: .. For RNase A or T1 digestion analysis, 70 pmol test RNA was incubated with RNase A (20 µg, Thermo Fisher Scientific) in 10 mM Tris-HCl pH < 7, 20 µM EDTA, 10 mM NaCl for 1 h at 60°C; or RNase T1 (3400 u, Thermo Fisher Scientific) in 50 mM Tris-HCl pH 6, 1 mM EDTA for 1 h at 57°C. .. Both reactions were further supplemented with phosphatase SAP (2 u, Fermentas) and incubated for 2 h at 37°C.

Article Title: Differential ratio amplicons (Ramp) for the evaluation of RNA integrity extracted from complex environmental samples
Article Snippet: .. Enzymatic degradation by RNase I For RNAse I degradation experiment, 40 μl aliquots of DNA‐free RNA was incubated at 37°C for 40 min in the presence of increasing concentrations of RNase I (Ambion): 0 (buffer only), 2, 10, 20 and 40 Units RNase I μg−1 RNA. .. The reaction was stopped by adding 10 μl β‐mercaptoethanol and RNA was recovered by ethanol precipitation: 5 μl of 7.5 M ammonium acetate and 137.5 μl 100% ethanol was added and the mixture was precipitated overnight at −20°C.

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: .. Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. Polynucleotides were removed from DNase I digestion products by using the QIAquick PCR Purification Kit (Qiagen).

Infection:

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. To test the cytotoxicity of cell culture supernatants infected with M. bovis , mycoplasma cells were killed by a gentamicin treatment (400 μg/ml) for 3 h at 37°C and mycoplasma cells were depleted by 30 min centrifugation at 12,000 ×g (4°C).

Expressing:

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: Paragraph title: Expression of reporters in the PURExpress in vitro translation system ... For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Article Title: Transcriptional coactivator PGC-1α contains a novel CBP80-binding motif that orchestrates efficient target gene expression
Article Snippet: Immunoprecipitations were performed after the addition of NaCl to 150 mM as described previously , except that RNase I (Invitrogen) was used in place of RNase A, and antibodies were as described for Western blotting (see below). .. For tandem immunoprecipitations, C2C12 cells expressing Flag-CBP80 were lysed and immunoprecipitated using anti-Flag M2 affinity beads (Sigma).

Western Blot:

Article Title: Transcriptional coactivator PGC-1α contains a novel CBP80-binding motif that orchestrates efficient target gene expression
Article Snippet: .. Immunoprecipitations were performed after the addition of NaCl to 150 mM as described previously , except that RNase I (Invitrogen) was used in place of RNase A, and antibodies were as described for Western blotting (see below). .. For tandem immunoprecipitations, C2C12 cells expressing Flag-CBP80 were lysed and immunoprecipitated using anti-Flag M2 affinity beads (Sigma).

Gel Purification:

Article Title: Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile
Article Snippet: After denaturing gel purification, 5’-32 P-labeled tRNA was mixed with non-radiolabeled tRNA and subjected to heat-denaturation at 65°C for 5 min in the respective reaction buffer suitable for partial RNase T1 or A digestion. .. Partial digestion of tRNA with RNase A: 1 μg of tRNA in 10 mM Tris–HCl, pH 7.5, and 1 mM EDTA was pre-incubated at 37°C for 5 min before RNase A (0.004 U; Ambion) was added and samples were incubated at 37°C for 5 min.

High Performance Liquid Chromatography:

Article Title: Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
Article Snippet: Paragraph title: HPLC-MS analysis ... For RNase A or T1 digestion analysis, 70 pmol test RNA was incubated with RNase A (20 µg, Thermo Fisher Scientific) in 10 mM Tris-HCl pH < 7, 20 µM EDTA, 10 mM NaCl for 1 h at 60°C; or RNase T1 (3400 u, Thermo Fisher Scientific) in 50 mM Tris-HCl pH 6, 1 mM EDTA for 1 h at 57°C.

Electron Microscopy:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Transfection:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: Reciprocal pulldown assays HEK293T cells that were cultured in 10-cm dish with 50–60% confluence were transfected with 8 μg of the FLAG-tagged wild-type or mutated ADAR1/2 plasmid, followed by the protein extraction 48 h post-transfection. .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C.

Concentration Assay:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: In every competition experiment, equal masses of the competitor and reference RNAs—each at a concentration of 30 ng/μl—were mixed in buffer B (BB; 1 M NaCl, 20 mM Tris-HCl [pH 7.2], 1 mM EDTA, 1 mM dithiothreitol [DTT], and 1 mM phenylmethanesulfonylfluoride [PMSF]), with dissociated CCMV CP subunits at a CP/total RNA ratio (wt/wt) of 3:1 (here total RNA is competitor RNA plus reference RNA). .. Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times.

Article Title: RNA-binding protein FXR1 is presented in rat brain in amyloid form
Article Snippet: .. Protein analysis Preparation of protein lysates was performed as described previously with addition of RNase A (Thermo Fisher, USA) to the lysis buffer to final concentration 200 µg/ml. ..

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: To test the influence of extracellular nucleic acids on M. bovis cytotoxicity, mycoplasmas and EBL cells were co-cultivated in DMEM-based medium supplemented with increasing concentration of calf thymus DNA (UltraPure™ Calf Thymus DNA Solution, Invitrogen) and/or yeast tRNA (Invitrogen). .. Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C.

Protease Inhibitor:

Article Title: Transcriptional coactivator PGC-1α contains a novel CBP80-binding motif that orchestrates efficient target gene expression
Article Snippet: Immunoprecipitations were performed after the addition of NaCl to 150 mM as described previously , except that RNase I (Invitrogen) was used in place of RNase A, and antibodies were as described for Western blotting (see below). .. Beads were washed twice using ice-cold NET-2 buffer (50 mM Tris-HCl at pH 7.4, 150 mM NaCl, 0.05% NP-40, Complete mini, EDTA-free protease inhibitor cocktail [Roche]).

Magnetic Beads:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C. .. In order to determine the binding of endogenous ADARs to Dicer, Huh-7 cells were analysed by reciprocal pulldown assays using the ADAR1, ADAR2 or Dicer-specific antibody.

Cell Culture:

Article Title: Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A
Article Snippet: Materials and Methods NIH/3T3 cells were cultured at 37°C, 5% CO2 in DMEM (Sigma) with 10% calf serum. .. RNase A (Roche 10109142001) was dissolved at 10 mg/ml in 10 mM sodium acetate pH 5.2, boiled for 15 min then neutralised by addition of Tris pH 7.4 to 0.1 M then diluted to a 1 mg/ml stock, 10 mg/ml DNase-free RNase (Thermo EN0531) was used as provided (0.1 µl per reaction).

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: Reciprocal pulldown assays HEK293T cells that were cultured in 10-cm dish with 50–60% confluence were transfected with 8 μg of the FLAG-tagged wild-type or mutated ADAR1/2 plasmid, followed by the protein extraction 48 h post-transfection. .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C.

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. To test the cytotoxicity of cell culture supernatants infected with M. bovis , mycoplasma cells were killed by a gentamicin treatment (400 μg/ml) for 3 h at 37°C and mycoplasma cells were depleted by 30 min centrifugation at 12,000 ×g (4°C).

Generated:

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: The peptidyl-tRNA construct was generated by creating a truncated mRNA lacking a stop codon directly after the Thrdx-HA sequence. .. For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Transmission Assay:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Sequencing:

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: The peptidyl-tRNA construct was generated by creating a truncated mRNA lacking a stop codon directly after the Thrdx-HA sequence. .. For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Binding Assay:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C. .. In order to determine the binding of endogenous ADARs to Dicer, Huh-7 cells were analysed by reciprocal pulldown assays using the ADAR1, ADAR2 or Dicer-specific antibody.

Immunofluorescence:

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death
Article Snippet: Immunofluorescence using Rae1 antibody was carried out as described by ). .. For RNase A or RNAsin pre-treatments, cell lysates were pre-incubated with RNAsin (1,000 units/ml) or RNase A (50 μg/ml; Ambion, Austin, TX, USA) for 15 min at 37°C, followed by incubation on ice for 20 min.

Transmission Electron Microscopy:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Nucleic Acid Electrophoresis:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Fluorescence:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: In the quantitative case, the RNAs are labeled with different fluorophores so that their relative numbers in RNase-treated assembly mixes can be assayed quantitatively by measuring the fluorescence spectra of the purified nucleocapsids. .. Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times.

Methylation:

Article Title: Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
Article Snippet: Methylation reactions for HPLC analysis typically contained 0.5–2 µM test RNA, 1–2 µM aFib–Nop5, and 100 µM SAM, if any. .. For RNase A or T1 digestion analysis, 70 pmol test RNA was incubated with RNase A (20 µg, Thermo Fisher Scientific) in 10 mM Tris-HCl pH < 7, 20 µM EDTA, 10 mM NaCl for 1 h at 60°C; or RNase T1 (3400 u, Thermo Fisher Scientific) in 50 mM Tris-HCl pH 6, 1 mM EDTA for 1 h at 57°C.

Isolation:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C. .. To conduct pulldown assays for different cell fractions, nuclear fractions, cytoplasmic fractions and total cell lysates were isolated from HEK293T cells, followed by reciprocal pulldown assays using the ADAR1, ADAR2 or Dicer-specific antibody.

Article Title: Novel Structure and Unexpected RNA-Binding Ability of the C-Terminal Domain of Herpes Simplex Virus 1 Tegument Protein UL21
Article Snippet: Total nucleic acids were isolated by phenol-chloroform precipitation. .. To isolate DNA only, the complex was treated with RNase A at an RNase A (Invitrogen)/UL21C volume ratio of 1:1,000 for 20 min at 37°C prior to nucleic acid extraction with the neutral pH phenol-chloroform mixture.

Microscopy:

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death
Article Snippet: Samples were analysed both by using the Metasystems (Boston, MA, USA) Zeiss Axioplan 2e with a Zeiss Axiocam HRm digital camera and a Leica (Bannockburn, IL, USA) TCS SP5 confocal microscope. .. For RNase A or RNAsin pre-treatments, cell lysates were pre-incubated with RNAsin (1,000 units/ml) or RNase A (50 μg/ml; Ambion, Austin, TX, USA) for 15 min at 37°C, followed by incubation on ice for 20 min.

Purification:

Article Title: FMRP recruitment of β‐catenin to the translation pre‐initiation complex represses translation
Article Snippet: Purified Wnt‐3a (315‐20) was purchased from PeproTech. .. RNase A was purchased from ThermoFisher (EN0531).

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm. .. To prepare the beads for RNA purification, 50 μl of protein G-coated Dynabeads (Invitrogen) was washed twice with 1 ml lysis buffer and resuspended in 200 μl lysis buffer containing 3–5 μg of RBFOX2 (Bethyl Laboratories, A300-864A) or CPSF2 (Bethyl Laboratories, A301-580A) antibody.

Article Title: Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile
Article Snippet: Analysis of 5’-32 P-labeled tRNA by partial RNase T1 and A digestion and alkali hydrolysis Purified tRNAs were dephosphorylated with calf intestinal alkaline phosphatase (NEB) and subsequently labeled at the 5′ terminus with 32 P using T4-PNK (NEB) following standard procedures ( ). .. Partial digestion of tRNA with RNase A: 1 μg of tRNA in 10 mM Tris–HCl, pH 7.5, and 1 mM EDTA was pre-incubated at 37°C for 5 min before RNase A (0.004 U; Ambion) was added and samples were incubated at 37°C for 5 min.

Article Title: Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A
Article Snippet: Major satellite PCR product was amplified using Taq polymerase (NEB) and primers T7 and T3 from pCR4-Maj9-2 , digested with Eco RI and gel purified using a QIAQuick column (QIAGEN). .. RNase A (Roche 10109142001) was dissolved at 10 mg/ml in 10 mM sodium acetate pH 5.2, boiled for 15 min then neutralised by addition of Tris pH 7.4 to 0.1 M then diluted to a 1 mg/ml stock, 10 mg/ml DNase-free RNase (Thermo EN0531) was used as provided (0.1 µl per reaction).

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: .. Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. Polynucleotides were removed from DNase I digestion products by using the QIAquick PCR Purification Kit (Qiagen).

Polymerase Chain Reaction:

Article Title: Unexpected DNA Loss Mediated by the DNA Binding Activity of Ribonuclease A
Article Snippet: Major satellite PCR product was amplified using Taq polymerase (NEB) and primers T7 and T3 from pCR4-Maj9-2 , digested with Eco RI and gel purified using a QIAQuick column (QIAGEN). .. RNase A (Roche 10109142001) was dissolved at 10 mg/ml in 10 mM sodium acetate pH 5.2, boiled for 15 min then neutralised by addition of Tris pH 7.4 to 0.1 M then diluted to a 1 mg/ml stock, 10 mg/ml DNase-free RNase (Thermo EN0531) was used as provided (0.1 µl per reaction).

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: The PURExpress reactions were initiated by mixing 1 μl of PCR product (29–22 ng/μl), 2 μl of solution A, 1.5 μl of solution B, and 0.6 μl of 35 S-methionine. .. For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C. .. Polynucleotides were removed from DNase I digestion products by using the QIAquick PCR Purification Kit (Qiagen).

Protein Extraction:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: Reciprocal pulldown assays HEK293T cells that were cultured in 10-cm dish with 50–60% confluence were transfected with 8 μg of the FLAG-tagged wild-type or mutated ADAR1/2 plasmid, followed by the protein extraction 48 h post-transfection. .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C.

Labeling:

Article Title: Protein-imprinted polysiloxane scaffolds
Article Snippet: .. Lysozyme was labeled with Alexa Fluor 488 (Molecular Probes; λex = 495 nm and λem = 519 nm), and RNase A was labeled with Alexa Fluor 594 (Molecular Probes; λex = 590 nm and λem = 617 nm). ..

Article Title: Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile
Article Snippet: Analysis of 5’-32 P-labeled tRNA by partial RNase T1 and A digestion and alkali hydrolysis Purified tRNAs were dephosphorylated with calf intestinal alkaline phosphatase (NEB) and subsequently labeled at the 5′ terminus with 32 P using T4-PNK (NEB) following standard procedures ( ). .. Partial digestion of tRNA with RNase A: 1 μg of tRNA in 10 mM Tris–HCl, pH 7.5, and 1 mM EDTA was pre-incubated at 37°C for 5 min before RNase A (0.004 U; Ambion) was added and samples were incubated at 37°C for 5 min.

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: In the quantitative case, the RNAs are labeled with different fluorophores so that their relative numbers in RNase-treated assembly mixes can be assayed quantitatively by measuring the fluorescence spectra of the purified nucleocapsids. .. Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times.

Lysis:

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: .. Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm. ..

Article Title: RNA-binding protein FXR1 is presented in rat brain in amyloid form
Article Snippet: .. Protein analysis Preparation of protein lysates was performed as described previously with addition of RNase A (Thermo Fisher, USA) to the lysis buffer to final concentration 200 µg/ml. ..

Titration:

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: At different time post-inoculation, mycoplasma titers were determined by CFU titration following one freeze-thaw cycle. .. Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C.

Plasmid Preparation:

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: Reciprocal pulldown assays HEK293T cells that were cultured in 10-cm dish with 50–60% confluence were transfected with 8 μg of the FLAG-tagged wild-type or mutated ADAR1/2 plasmid, followed by the protein extraction 48 h post-transfection. .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C.

Irradiation:

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: Flp-In-293 cells were grown in 10 cm plates and then subjected to UV-C irradiation (200 mJ/cm2 , Stratalinker 2400). .. Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm.

In Vitro:

Article Title: Ribosomes slide on lysine-encoding homopolymeric A stretches
Article Snippet: Paragraph title: Expression of reporters in the PURExpress in vitro translation system ... For the experiments in which the PURExpress reaction products were treated with RNase A , 0.5–1 μg of RNase A (Ambion, Grand Island, NY) was added to each reaction and solutions were incubated on ice for an additional 30 min before being denatured and loaded on a gel.

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Paragraph title: In vitro assemblies for the qualitative competition experiments. ... Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times.

Protein Binding:

Article Title: Protein-imprinted polysiloxane scaffolds
Article Snippet: Paragraph title: 2.4. Preferential protein binding ... Lysozyme was labeled with Alexa Fluor 488 (Molecular Probes; λex = 495 nm and λem = 519 nm), and RNase A was labeled with Alexa Fluor 594 (Molecular Probes; λex = 590 nm and λem = 617 nm).

Immunoprecipitation:

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death
Article Snippet: Immunoprecipitation assays. .. For RNase A or RNAsin pre-treatments, cell lysates were pre-incubated with RNAsin (1,000 units/ml) or RNase A (50 μg/ml; Ambion, Austin, TX, USA) for 15 min at 37°C, followed by incubation on ice for 20 min.

Article Title: Global analysis of CPSF2-mediated alternative splicing: Integration of global iCLIP and transcriptome profiling data
Article Snippet: Paragraph title: Individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) ... Samples were partially digested with RNase I by incubation with 10 μl RNase I (Ambion) (diluted 1:500 in lysis buffer) and 5 μl Turbo DNase (Ambion) for 5 min at 37 °C while shaking at 1100 rpm.

Article Title: An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer
Article Snippet: .. For the treatment of RNase A prior to the immunoprecipitation, the total lysates were incubated with 1 μg/ml RNase A (20 mg/ml, Thermo Fisher Scientific, MA, USA) at 37 °C for 30 min. To pulldown FLAG-tagged protein, ∼5 mg of total cell lysates were immunoprecipitated with anti-FLAG M2 Magnetic Beads (Sigma-Aldrich) for 2 h at 4°C. .. In order to determine the binding of endogenous ADARs to Dicer, Huh-7 cells were analysed by reciprocal pulldown assays using the ADAR1, ADAR2 or Dicer-specific antibody.

Article Title: Transcriptional coactivator PGC-1α contains a novel CBP80-binding motif that orchestrates efficient target gene expression
Article Snippet: Immunoprecipitations were performed after the addition of NaCl to 150 mM as described previously , except that RNase I (Invitrogen) was used in place of RNase A, and antibodies were as described for Western blotting (see below). .. For tandem immunoprecipitations, C2C12 cells expressing Flag-CBP80 were lysed and immunoprecipitated using anti-Flag M2 affinity beads (Sigma).

Ethanol Precipitation:

Article Title: Differential ratio amplicons (Ramp) for the evaluation of RNA integrity extracted from complex environmental samples
Article Snippet: Enzymatic degradation by RNase I For RNAse I degradation experiment, 40 μl aliquots of DNA‐free RNA was incubated at 37°C for 40 min in the presence of increasing concentrations of RNase I (Ambion): 0 (buffer only), 2, 10, 20 and 40 Units RNase I μg−1 RNA. .. The reaction was stopped by adding 10 μl β‐mercaptoethanol and RNA was recovered by ethanol precipitation: 5 μl of 7.5 M ammonium acetate and 137.5 μl 100% ethanol was added and the mixture was precipitated overnight at −20°C.

Staining:

Article Title: Extracellular DNA: A Nutritional Trigger of Mycoplasma bovis Cytotoxicity
Article Snippet: Cell monolayers were washed with Dulbecco’s phosphate-buffered saline (DPBS, Gibco) and stained for 60 min with 0.1% crystal violet solution in 10% ethanol. .. Calf thymus DNA treatment with Proteinase K (Qiagen), DNase I (Invitrogen), and RNase A (Invitrogen) was performed by 1 h incubation in DMEM medium and 15 min heating at 96°C.

UV-Vis Spectroscopy:

Article Title: In Vitro Quantification of the Relative Packaging Efficiencies of Single-Stranded RNA Molecules by Viral Capsid Protein
Article Snippet: Aliquots (8 μl) of each sample and control were set aside for electrophoretic mobility assays, while the rest was treated with RNase A (Invitrogen) (4 ng/μl) for 45 min. RNase A was inactivated by incubating the samples with RNase inhibitor (Roche) for 15 min. All samples and controls were purified and concentrated by washing with VSB using a 100-kDa Amicon centrifuge filter (0.5 ml; Millipore) at 3,000 × g for 5 min; this step was repeated 4 times. .. Two 6-μl aliquots of each sample and control were set aside for UV-visible (UV-Vis) spectroscopy and transmission electron microscopy (TEM), and the rest was used to analyze the RNA content by gel electrophoresis.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Thermo Fisher rnase t1
    Rnase T1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase t1/product/Thermo Fisher
    Average 90 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    rnase t1 - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    90
    Thermo Fisher ribolock rnase inhibitor
    Ribolock Rnase Inhibitor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ribolock rnase inhibitor/product/Thermo Fisher
    Average 90 stars, based on 90 article reviews
    Price from $9.99 to $1999.99
    ribolock rnase inhibitor - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier


    Image Search Results