nascent viral rna  (Thermo Fisher)


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    Structured Review

    Thermo Fisher nascent viral rna
    Location of viral <t>RNA</t> and protein 2C in the infected cell. <t>HPEV-1-infected</t> A549 cells, harvested at 4 (a and e), 6 (b and f), 8 (c and g) and 10 (d and h) h p.i., were subjected to FISH with an FITC-coupled HPEV plus-strand specific riboprobe (a to d) or to IF with Ab against protein 2C (e to h). Bar, 25 μm.
    Nascent Viral Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 43832 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nascent viral rna/product/Thermo Fisher
    Average 85 stars, based on 43832 article reviews
    Price from $9.99 to $1999.99
    nascent viral rna - by Bioz Stars, 2021-01
    85/100 stars

    Images

    1) Product Images from "Replication Complex of Human Parechovirus 1"

    Article Title: Replication Complex of Human Parechovirus 1

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.15.8512-8523.2003

    Location of viral RNA and protein 2C in the infected cell. HPEV-1-infected A549 cells, harvested at 4 (a and e), 6 (b and f), 8 (c and g) and 10 (d and h) h p.i., were subjected to FISH with an FITC-coupled HPEV plus-strand specific riboprobe (a to d) or to IF with Ab against protein 2C (e to h). Bar, 25 μm.
    Figure Legend Snippet: Location of viral RNA and protein 2C in the infected cell. HPEV-1-infected A549 cells, harvested at 4 (a and e), 6 (b and f), 8 (c and g) and 10 (d and h) h p.i., were subjected to FISH with an FITC-coupled HPEV plus-strand specific riboprobe (a to d) or to IF with Ab against protein 2C (e to h). Bar, 25 μm.

    Techniques Used: Infection, Fluorescence In Situ Hybridization

    Association of HPEV-1 RNA with small vesicles (arrowheads), shown by EM-ISH with a DIG-labeled riboprobe immunodetected with an anti-DIG Ab and 10-nm-diameter gold-conjugated secondary Ab. The inset shows an example of a cluster of vesicles. Bars, 0.2 μm.
    Figure Legend Snippet: Association of HPEV-1 RNA with small vesicles (arrowheads), shown by EM-ISH with a DIG-labeled riboprobe immunodetected with an anti-DIG Ab and 10-nm-diameter gold-conjugated secondary Ab. The inset shows an example of a cluster of vesicles. Bars, 0.2 μm.

    Techniques Used: In Situ Hybridization, Labeling

    Colocalization of nascent viral RNA with protein 2C or the trans -Golgi marker GalT in HPEV-1-infected, Br-UTP-transfected A549 cells at 6 h p.i. (a) Br-RNA (green) detected with an Ab against Br-dUTP and Alexa 488-conjugated secondary Ab. (b) Protein 2C (red) detected with an Ab against 2C and Texas Red-conjugated secondary Ab. (c) Merge of panels a and b. (d) Br-RNA (red) detected with an Ab against Br-dUTP and Texas Red-conjugated secondary Ab. (e) GalT (green) visualized with an anti-GalT Ab and FITC-conjugated secondary Ab. (f) Merge of panels d and e. Picture sizes, 25 by 25 μm.
    Figure Legend Snippet: Colocalization of nascent viral RNA with protein 2C or the trans -Golgi marker GalT in HPEV-1-infected, Br-UTP-transfected A549 cells at 6 h p.i. (a) Br-RNA (green) detected with an Ab against Br-dUTP and Alexa 488-conjugated secondary Ab. (b) Protein 2C (red) detected with an Ab against 2C and Texas Red-conjugated secondary Ab. (c) Merge of panels a and b. (d) Br-RNA (red) detected with an Ab against Br-dUTP and Texas Red-conjugated secondary Ab. (e) GalT (green) visualized with an anti-GalT Ab and FITC-conjugated secondary Ab. (f) Merge of panels d and e. Picture sizes, 25 by 25 μm.

    Techniques Used: Marker, Infection, Transfection

    Characterization of viral macromolecules in HPEV-1-infected cells. (A) Kinetics of viral RNA synthesis in infected cells determined by [ 3 H]uridine labeling of nascent viral RNA. DPM, disintegrations per minute. (B) Immunoblots of mock-infected (lane 1) and HPEV-1-infected (lanes 2 to 4) cells (harvested at 4, 6, and 8 h p.i., respectively) were probed with Ab against protein 2C. Protein 2C accumulated in the absence of the precursor protein 2BC. (C) In vitro-translated proteins 2C (lane 1) and 2BC (lane 2) were immunoprecipitated with 2C antiserum and electrophoresed. The autoradiograph of the blot indicates that the 2C Ab was capable of recognizing 2C as well as the precursor protein 2BC.
    Figure Legend Snippet: Characterization of viral macromolecules in HPEV-1-infected cells. (A) Kinetics of viral RNA synthesis in infected cells determined by [ 3 H]uridine labeling of nascent viral RNA. DPM, disintegrations per minute. (B) Immunoblots of mock-infected (lane 1) and HPEV-1-infected (lanes 2 to 4) cells (harvested at 4, 6, and 8 h p.i., respectively) were probed with Ab against protein 2C. Protein 2C accumulated in the absence of the precursor protein 2BC. (C) In vitro-translated proteins 2C (lane 1) and 2BC (lane 2) were immunoprecipitated with 2C antiserum and electrophoresed. The autoradiograph of the blot indicates that the 2C Ab was capable of recognizing 2C as well as the precursor protein 2BC.

    Techniques Used: Infection, Labeling, Western Blot, In Vitro, Immunoprecipitation, Autoradiography

    2) Product Images from "Replication Complex of Human Parechovirus 1"

    Article Title: Replication Complex of Human Parechovirus 1

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.15.8512-8523.2003

    Location of protein 2C visualized by confocal scanning laser microscopy. (a) Double labeling of infected cell for protein 2C and Golgi protein GM130 shows the disintegration of the Golgi apparatus in the infected cell. Protein 2C was visualized with Texas Red-conjugated secondary Ab (red), and GM130 was visualized with Alexa 488-conjugated secondary Ab (green). 1, uninfected cell with intact Golgi; 2, HPEV-infected cell with dispersed Golgi complex. (b to f) Optical sectioning through an HPEV-1-infected cell 6 h p.i., stained with Ab against protein 2C. The distance between two consecutive sections is 0.5 μm. Protein 2C is located on branching, stick-like structures. (g to i) Double labeling of infected cells with Abs against protein 2C (red) and the cytoskeleton markers (green) tubulin (g) and vimentin (h) and with Alexa 488-coupled phalloidin that labels actin filaments (i). 1, uninfected cell with intact actin filaments; 2, HPEV-infected cell with depolymerized actin filaments. Picture sizes are as follows: a, 44 by 44 μm; b to f, 25 by 25 μm; g, 19 by 19 μm; h, 30 by 30 μm; i, 50 by 50 μm.
    Figure Legend Snippet: Location of protein 2C visualized by confocal scanning laser microscopy. (a) Double labeling of infected cell for protein 2C and Golgi protein GM130 shows the disintegration of the Golgi apparatus in the infected cell. Protein 2C was visualized with Texas Red-conjugated secondary Ab (red), and GM130 was visualized with Alexa 488-conjugated secondary Ab (green). 1, uninfected cell with intact Golgi; 2, HPEV-infected cell with dispersed Golgi complex. (b to f) Optical sectioning through an HPEV-1-infected cell 6 h p.i., stained with Ab against protein 2C. The distance between two consecutive sections is 0.5 μm. Protein 2C is located on branching, stick-like structures. (g to i) Double labeling of infected cells with Abs against protein 2C (red) and the cytoskeleton markers (green) tubulin (g) and vimentin (h) and with Alexa 488-coupled phalloidin that labels actin filaments (i). 1, uninfected cell with intact actin filaments; 2, HPEV-infected cell with depolymerized actin filaments. Picture sizes are as follows: a, 44 by 44 μm; b to f, 25 by 25 μm; g, 19 by 19 μm; h, 30 by 30 μm; i, 50 by 50 μm.

    Techniques Used: Microscopy, Labeling, Infection, Staining

    Location of viral RNA and protein 2C in the infected cell. HPEV-1-infected A549 cells, harvested at 4 (a and e), 6 (b and f), 8 (c and g) and 10 (d and h) h p.i., were subjected to FISH with an FITC-coupled HPEV plus-strand specific riboprobe (a to d) or to IF with Ab against protein 2C (e to h). Bar, 25 μm.
    Figure Legend Snippet: Location of viral RNA and protein 2C in the infected cell. HPEV-1-infected A549 cells, harvested at 4 (a and e), 6 (b and f), 8 (c and g) and 10 (d and h) h p.i., were subjected to FISH with an FITC-coupled HPEV plus-strand specific riboprobe (a to d) or to IF with Ab against protein 2C (e to h). Bar, 25 μm.

    Techniques Used: Infection, Fluorescence In Situ Hybridization

    Association of HPEV-1 RNA with small vesicles (arrowheads), shown by EM-ISH with a DIG-labeled riboprobe immunodetected with an anti-DIG Ab and 10-nm-diameter gold-conjugated secondary Ab. The inset shows an example of a cluster of vesicles. Bars, 0.2 μm.
    Figure Legend Snippet: Association of HPEV-1 RNA with small vesicles (arrowheads), shown by EM-ISH with a DIG-labeled riboprobe immunodetected with an anti-DIG Ab and 10-nm-diameter gold-conjugated secondary Ab. The inset shows an example of a cluster of vesicles. Bars, 0.2 μm.

    Techniques Used: In Situ Hybridization, Labeling

    Colocalization of nascent viral RNA with protein 2C or the trans -Golgi marker GalT in HPEV-1-infected, Br-UTP-transfected A549 cells at 6 h p.i. (a) Br-RNA (green) detected with an Ab against Br-dUTP and Alexa 488-conjugated secondary Ab. (b) Protein 2C (red) detected with an Ab against 2C and Texas Red-conjugated secondary Ab. (c) Merge of panels a and b. (d) Br-RNA (red) detected with an Ab against Br-dUTP and Texas Red-conjugated secondary Ab. (e) GalT (green) visualized with an anti-GalT Ab and FITC-conjugated secondary Ab. (f) Merge of panels d and e. Picture sizes, 25 by 25 μm.
    Figure Legend Snippet: Colocalization of nascent viral RNA with protein 2C or the trans -Golgi marker GalT in HPEV-1-infected, Br-UTP-transfected A549 cells at 6 h p.i. (a) Br-RNA (green) detected with an Ab against Br-dUTP and Alexa 488-conjugated secondary Ab. (b) Protein 2C (red) detected with an Ab against 2C and Texas Red-conjugated secondary Ab. (c) Merge of panels a and b. (d) Br-RNA (red) detected with an Ab against Br-dUTP and Texas Red-conjugated secondary Ab. (e) GalT (green) visualized with an anti-GalT Ab and FITC-conjugated secondary Ab. (f) Merge of panels d and e. Picture sizes, 25 by 25 μm.

    Techniques Used: Marker, Infection, Transfection

    Characterization of viral macromolecules in HPEV-1-infected cells. (A) Kinetics of viral RNA synthesis in infected cells determined by [ 3 H]uridine labeling of nascent viral RNA. DPM, disintegrations per minute. (B) Immunoblots of mock-infected (lane 1) and HPEV-1-infected (lanes 2 to 4) cells (harvested at 4, 6, and 8 h p.i., respectively) were probed with Ab against protein 2C. Protein 2C accumulated in the absence of the precursor protein 2BC. (C) In vitro-translated proteins 2C (lane 1) and 2BC (lane 2) were immunoprecipitated with 2C antiserum and electrophoresed. The autoradiograph of the blot indicates that the 2C Ab was capable of recognizing 2C as well as the precursor protein 2BC.
    Figure Legend Snippet: Characterization of viral macromolecules in HPEV-1-infected cells. (A) Kinetics of viral RNA synthesis in infected cells determined by [ 3 H]uridine labeling of nascent viral RNA. DPM, disintegrations per minute. (B) Immunoblots of mock-infected (lane 1) and HPEV-1-infected (lanes 2 to 4) cells (harvested at 4, 6, and 8 h p.i., respectively) were probed with Ab against protein 2C. Protein 2C accumulated in the absence of the precursor protein 2BC. (C) In vitro-translated proteins 2C (lane 1) and 2BC (lane 2) were immunoprecipitated with 2C antiserum and electrophoresed. The autoradiograph of the blot indicates that the 2C Ab was capable of recognizing 2C as well as the precursor protein 2BC.

    Techniques Used: Infection, Labeling, Western Blot, In Vitro, Immunoprecipitation, Autoradiography

    Sequence comparison of human picornavirus 2C protein. (A) Phylogenetic tree of human picornaviruses based on the 2C genes. HRV, human rhinovirus; CAV, coxsackie A virus; CBV, coxsackie B virus. (B) Sequence alignment of the 2C proteins of HPEV-1, PV, and HAV. The three ATPase motifs are boxed, and 179N in the PV sequence and the corresponding glycine residues in the HPEV-1 and HAV sequences as well as 187M in the PV sequence and its corresponding amino acids in the HPEV-1 and HAV sequences, are highlighted. The cysteine-rich region in the PV sequence is underlined.
    Figure Legend Snippet: Sequence comparison of human picornavirus 2C protein. (A) Phylogenetic tree of human picornaviruses based on the 2C genes. HRV, human rhinovirus; CAV, coxsackie A virus; CBV, coxsackie B virus. (B) Sequence alignment of the 2C proteins of HPEV-1, PV, and HAV. The three ATPase motifs are boxed, and 179N in the PV sequence and the corresponding glycine residues in the HPEV-1 and HAV sequences as well as 187M in the PV sequence and its corresponding amino acids in the HPEV-1 and HAV sequences, are highlighted. The cysteine-rich region in the PV sequence is underlined.

    Techniques Used: Sequencing

    Electron micrographs of A549 cells demonstrate structural changes seen 6 h after HPEV-1 infection. (a) Uninfected cell; (b) infected cell, dashed line delineates region with alterations in the cellular architecture; (c and d) higher magnification shows the main features of cell alterations, including the appearance of clusters of small vesicles (V) (c), dilated ER (arrowheads), and rough ER (rER) (d). N, nucleus. Bars, 0.5 μm.
    Figure Legend Snippet: Electron micrographs of A549 cells demonstrate structural changes seen 6 h after HPEV-1 infection. (a) Uninfected cell; (b) infected cell, dashed line delineates region with alterations in the cellular architecture; (c and d) higher magnification shows the main features of cell alterations, including the appearance of clusters of small vesicles (V) (c), dilated ER (arrowheads), and rough ER (rER) (d). N, nucleus. Bars, 0.5 μm.

    Techniques Used: Infection

    Related Articles

    Immunohistochemistry:

    Article Title: Cdc6 Chromatin Affinity Is Unaffected by Serine-54 Phosphorylation, S-Phase Progression, and Overexpression of Cyclin A
    Article Snippet: .. Monoclonal antihemagglutinin (anti-HA), nonconjugated or conjugated with fluorescein isothiocyanate (FITC) (1:1,000; HA.11; Covance), and rabbit polyclonal anti-Cdc6 (sc-8341, antibody 1) and anti-Cdc6-phosphoserine-54 (sc-12920; 1:500) and blocking peptide (sc-12920p) were from Santa Cruz Biotechnology; rabbit polyclonal anti-Mcm5 (1:50; provided by Rolf Knippers, University of Konstanz, Konstanz, Germany), monoclonal anti-BrdU (1:20; Roche), monoclonal anti-Cdc6 37314 (against amino acids 366 to 560 of human Cdc6; antibody 2; 1:1,000), and monoclonal anti-Cdc6 16724 (against amino acids 1 to 224 of human Cdc6; antibody 3; 1:1,000 on immunoblots and 1:500 in immunohistochemistry) were provided by Nicholas Heintz (University of Vermont); monoclonal anti-BrdU-Alexa-594 (1:20) and monoclonal anti-Cdc6 (antibody 4; immunogenic peptide shown in Fig. ; 1:20) were from Molecular Probes; monoclonal anti-lamin A/C (1:500) and monoclonal antitubulin (1:1,000) were from Calbiochem; rabbit polyclonal anti-cyclin A (1 μg/ml) was from Upstate Biotechnology Inc. .. Human and murine Cdc6 sequences (GenBank accession numbers and NM_ 011799 , respectively) were aligned, and identical stretches of DNA sequences were used to design primers for use in 5′ rapid amplification of cDNA ends (5′-RACE) and reverse transcription-PCRs (RT-PCRs) on Chinese hamster cDNA.

    BrdU Incorporation Assay:

    Article Title: Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch
    Article Snippet: .. EdU incorporation was detected using the Click-iT EdU Alexa Fluor 488 Flow Cytometry Assay Kit (Invitrogen C10425), and BrdU incorporation was detected using Alexa Fluor 647 mouse anti-BrdU (Invitrogen B35133). .. In vitro CFUe expansion cultures ( , ) To isolate Ter119-negative cells, fresh fetal liver cells were stained with biotin-conjugated anti-Ter119 (BD Biosciences 553672) at a 1:100 ratio, followed by EasySep magnetic separation (StemCell Technologies).

    Labeling:

    Article Title: Discrimination Between Lung Homeostatic and Injury-Induced Epithelial Progenitor Subsets by Cell-Density Properties
    Article Snippet: .. Fractionated cells were washed and labeled with the following antibodies: FITC-conjugated anti-CD45R (Invitrogen), R-PE anti- SCA-1 (Ly6A/E; Invitrogen), PE/Cy5 anti- c-KIT (CD117; eBioscience), PE-anti-CD49f (eBioscience), Alexa-488-anti-EpCAM (CD326; Biolegend, San Diego, CA), biotinylated-CD31 (Santa Cruz Biotech) followed by PE/Cy5 streptavidin (Biolegend), and mouse anti-BRDU (ebioscience) followed by anti-mouse Alexa 488 (Invitrogen). .. Isotype controls included Alexa Fluor® 488-conjugated rat-IgG, FITC-rat IgG, R-PE-rabbit IgG (Southern Biotech, Birmingham, AL), and PE/Cy5 IgG (BD Pharmingen, San Diego, CA).

    Immunolabeling:

    Article Title: Modulation of subventricular zone oligodendrogenesis: a role for hemopressin?
    Article Snippet: .. Then, SVZ cells were fixed in PFA and BrdU was immunolabeled with the anti-BrdU mouse monoclonal antibody (Clone MoBU-1) Alexa Fluor 594 (Invitrogen) as described previously ( ; Figure ). ..

    Cytometry:

    Article Title: Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch
    Article Snippet: .. EdU incorporation was detected using the Click-iT EdU Alexa Fluor 488 Flow Cytometry Assay Kit (Invitrogen C10425), and BrdU incorporation was detected using Alexa Fluor 647 mouse anti-BrdU (Invitrogen B35133). .. In vitro CFUe expansion cultures ( , ) To isolate Ter119-negative cells, fresh fetal liver cells were stained with biotin-conjugated anti-Ter119 (BD Biosciences 553672) at a 1:100 ratio, followed by EasySep magnetic separation (StemCell Technologies).

    Blocking Assay:

    Article Title: Cdc6 Chromatin Affinity Is Unaffected by Serine-54 Phosphorylation, S-Phase Progression, and Overexpression of Cyclin A
    Article Snippet: .. Monoclonal antihemagglutinin (anti-HA), nonconjugated or conjugated with fluorescein isothiocyanate (FITC) (1:1,000; HA.11; Covance), and rabbit polyclonal anti-Cdc6 (sc-8341, antibody 1) and anti-Cdc6-phosphoserine-54 (sc-12920; 1:500) and blocking peptide (sc-12920p) were from Santa Cruz Biotechnology; rabbit polyclonal anti-Mcm5 (1:50; provided by Rolf Knippers, University of Konstanz, Konstanz, Germany), monoclonal anti-BrdU (1:20; Roche), monoclonal anti-Cdc6 37314 (against amino acids 366 to 560 of human Cdc6; antibody 2; 1:1,000), and monoclonal anti-Cdc6 16724 (against amino acids 1 to 224 of human Cdc6; antibody 3; 1:1,000 on immunoblots and 1:500 in immunohistochemistry) were provided by Nicholas Heintz (University of Vermont); monoclonal anti-BrdU-Alexa-594 (1:20) and monoclonal anti-Cdc6 (antibody 4; immunogenic peptide shown in Fig. ; 1:20) were from Molecular Probes; monoclonal anti-lamin A/C (1:500) and monoclonal antitubulin (1:1,000) were from Calbiochem; rabbit polyclonal anti-cyclin A (1 μg/ml) was from Upstate Biotechnology Inc. .. Human and murine Cdc6 sequences (GenBank accession numbers and NM_ 011799 , respectively) were aligned, and identical stretches of DNA sequences were used to design primers for use in 5′ rapid amplification of cDNA ends (5′-RACE) and reverse transcription-PCRs (RT-PCRs) on Chinese hamster cDNA.

    Western Blot:

    Article Title: Cdc6 Chromatin Affinity Is Unaffected by Serine-54 Phosphorylation, S-Phase Progression, and Overexpression of Cyclin A
    Article Snippet: .. Monoclonal antihemagglutinin (anti-HA), nonconjugated or conjugated with fluorescein isothiocyanate (FITC) (1:1,000; HA.11; Covance), and rabbit polyclonal anti-Cdc6 (sc-8341, antibody 1) and anti-Cdc6-phosphoserine-54 (sc-12920; 1:500) and blocking peptide (sc-12920p) were from Santa Cruz Biotechnology; rabbit polyclonal anti-Mcm5 (1:50; provided by Rolf Knippers, University of Konstanz, Konstanz, Germany), monoclonal anti-BrdU (1:20; Roche), monoclonal anti-Cdc6 37314 (against amino acids 366 to 560 of human Cdc6; antibody 2; 1:1,000), and monoclonal anti-Cdc6 16724 (against amino acids 1 to 224 of human Cdc6; antibody 3; 1:1,000 on immunoblots and 1:500 in immunohistochemistry) were provided by Nicholas Heintz (University of Vermont); monoclonal anti-BrdU-Alexa-594 (1:20) and monoclonal anti-Cdc6 (antibody 4; immunogenic peptide shown in Fig. ; 1:20) were from Molecular Probes; monoclonal anti-lamin A/C (1:500) and monoclonal antitubulin (1:1,000) were from Calbiochem; rabbit polyclonal anti-cyclin A (1 μg/ml) was from Upstate Biotechnology Inc. .. Human and murine Cdc6 sequences (GenBank accession numbers and NM_ 011799 , respectively) were aligned, and identical stretches of DNA sequences were used to design primers for use in 5′ rapid amplification of cDNA ends (5′-RACE) and reverse transcription-PCRs (RT-PCRs) on Chinese hamster cDNA.

    Flow Cytometry:

    Article Title: Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch
    Article Snippet: .. EdU incorporation was detected using the Click-iT EdU Alexa Fluor 488 Flow Cytometry Assay Kit (Invitrogen C10425), and BrdU incorporation was detected using Alexa Fluor 647 mouse anti-BrdU (Invitrogen B35133). .. In vitro CFUe expansion cultures ( , ) To isolate Ter119-negative cells, fresh fetal liver cells were stained with biotin-conjugated anti-Ter119 (BD Biosciences 553672) at a 1:100 ratio, followed by EasySep magnetic separation (StemCell Technologies).

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