pcrii topo vector  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pcrii topo vector
    Schematic presentation of steps for construction of the luciferase expression vector for transfection in L. donovani promastigotes. Partial maps with the relevant restriction sites are shown. (A) <t>pCRII-TOPO-</t> luc construct digested with SpeI and (B) pKS-Neo- luc clones digested with BamHI and EcoRI for determination of orientation of the luc ORF within the construct.
    Pcrii Topo Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcrii topo vector/product/Thermo Fisher
    Average 94 stars, based on 800 article reviews
    Price from $9.99 to $1999.99
    pcrii topo vector - by Bioz Stars, 2020-08
    94/100 stars

    Images

    1) Product Images from "Use of Leishmania donovani Field Isolates Expressing the Luciferase Reporter Gene in In Vitro Drug Screening †"

    Article Title: Use of Leishmania donovani Field Isolates Expressing the Luciferase Reporter Gene in In Vitro Drug Screening †

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.49.9.3776-3783.2005

    Schematic presentation of steps for construction of the luciferase expression vector for transfection in L. donovani promastigotes. Partial maps with the relevant restriction sites are shown. (A) pCRII-TOPO- luc construct digested with SpeI and (B) pKS-Neo- luc clones digested with BamHI and EcoRI for determination of orientation of the luc ORF within the construct.
    Figure Legend Snippet: Schematic presentation of steps for construction of the luciferase expression vector for transfection in L. donovani promastigotes. Partial maps with the relevant restriction sites are shown. (A) pCRII-TOPO- luc construct digested with SpeI and (B) pKS-Neo- luc clones digested with BamHI and EcoRI for determination of orientation of the luc ORF within the construct.

    Techniques Used: Luciferase, Expressing, Plasmid Preparation, Transfection, Construct, Clone Assay

    2) Product Images from "Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target"

    Article Title: Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target

    Journal: PLoS Medicine

    doi: 10.1371/journal.pmed.0040206

    Survival of Schistosomula after RNAi Silencing of TGR Expression (A) Qualitative measure of TGR transcripts by reverse transcription PCR. The abundance of TGR mRNA was greatly reduced by dsRNA treatment while the control gene GAPDH was unaffected. (B) Parasites were cultured in the presence of double-stranded TGR RNA (dashed lines) or of double-stranded irrelevant RNA (solid lines) in 20% O 2 (♦) or 0% O 2 (). A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR cloned into PCRII-TOPO vector was used to transcribe TGR dsRNA. Irrelevant, nonschistosome dsRNA used for negative controls was synthesized from the PCRII-TOPO vector using T7 and SP6 RNA polymerases. (C) Combination of RNAi and drug treatments. Solid lines are irrelevant dsRNA treatments and dashed lines are dsTGR treatments; no additions (♦), 2 μM praziquantel () and 2 μM AF (▴). At each time point, data comparing the combination treatment (2 μM AF + TGR dsRNA) to either 2 μM AF or TGR dsRNA alone were statistically significant (* p
    Figure Legend Snippet: Survival of Schistosomula after RNAi Silencing of TGR Expression (A) Qualitative measure of TGR transcripts by reverse transcription PCR. The abundance of TGR mRNA was greatly reduced by dsRNA treatment while the control gene GAPDH was unaffected. (B) Parasites were cultured in the presence of double-stranded TGR RNA (dashed lines) or of double-stranded irrelevant RNA (solid lines) in 20% O 2 (♦) or 0% O 2 (). A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR cloned into PCRII-TOPO vector was used to transcribe TGR dsRNA. Irrelevant, nonschistosome dsRNA used for negative controls was synthesized from the PCRII-TOPO vector using T7 and SP6 RNA polymerases. (C) Combination of RNAi and drug treatments. Solid lines are irrelevant dsRNA treatments and dashed lines are dsTGR treatments; no additions (♦), 2 μM praziquantel () and 2 μM AF (▴). At each time point, data comparing the combination treatment (2 μM AF + TGR dsRNA) to either 2 μM AF or TGR dsRNA alone were statistically significant (* p

    Techniques Used: Expressing, Polymerase Chain Reaction, Cell Culture, Clone Assay, Plasmid Preparation, Synthesized

    3) Product Images from "The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation"

    Article Title: The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw667

    Transcription of SARS mRNA starts at the annotated proximal promoter. Human hepatocellular carcinoma (HepG2) cells were incubated for 8 h in DMEM or DMEM + 5 mM HisOH to activate the AAR. The total RNA was subjected to RLM-5′ RACE analysis, as described in the Materials and Methods section. PCR products amplified with a 5′ RACE primer and a gene-specific primer were cloned into the pCRII-TOPO TA vector and the resulting clones were randomly selected for sequencing analysis. The top panel is the SARS gene structure; the grey boxes are the first three exons and the dotted oval is the approximate location of the CARE element. The arrow indicates the annotated transcription start site. The gene-specific primer used for PCR is labeled with a thick line under Exon 3. In the lower panels showing sequence, the annotated transcription start site is indicated with a ‘+1’ and the bold ATG represents the translation start codon. The underlined sequence is the location of the gene-specific primer. The multiple arrows inside the sequences are the actual transcription start sites established by the analysis of the clones obtained by the RLM-5′ RACE experiments.
    Figure Legend Snippet: Transcription of SARS mRNA starts at the annotated proximal promoter. Human hepatocellular carcinoma (HepG2) cells were incubated for 8 h in DMEM or DMEM + 5 mM HisOH to activate the AAR. The total RNA was subjected to RLM-5′ RACE analysis, as described in the Materials and Methods section. PCR products amplified with a 5′ RACE primer and a gene-specific primer were cloned into the pCRII-TOPO TA vector and the resulting clones were randomly selected for sequencing analysis. The top panel is the SARS gene structure; the grey boxes are the first three exons and the dotted oval is the approximate location of the CARE element. The arrow indicates the annotated transcription start site. The gene-specific primer used for PCR is labeled with a thick line under Exon 3. In the lower panels showing sequence, the annotated transcription start site is indicated with a ‘+1’ and the bold ATG represents the translation start codon. The underlined sequence is the location of the gene-specific primer. The multiple arrows inside the sequences are the actual transcription start sites established by the analysis of the clones obtained by the RLM-5′ RACE experiments.

    Techniques Used: Incubation, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation, Sequencing, Labeling

    4) Product Images from "Lrrc10 is required for early heart development and function in zebrafish"

    Article Title: Lrrc10 is required for early heart development and function in zebrafish

    Journal: Developmental biology

    doi: 10.1016/j.ydbio.2007.06.005

    Cardiac-specific expression of zlrrc10 . (A) The genomic structure of zlrrc10 . The full-length zlrrc10 cDNA consists of two exons. The size of exons, intron and the 5′ and 3′ UTR regions are shown in bp. The amino acid coding regions are represented by the hatched box. (B) An alignment of zebrafish, mouse and human LRRC10 amino acid sequences. Identical residues between all three species are shaded black, and the residues with similar characteristics are shaded gray. The conserved seven LRR motifs are numbered. (C) Cardiac-specific expression of lrrc10 in zebrafish embryos. Zebrafish embryos at 1 dpf (a–c) and 2 dpf (d–f) were subjected to whole-mount in situ hybridization using digoxigenin labeled zlrrc10 antisense cRNA probes (a, d). cmlc2 antisense cRNA probe (b and e) was used for positive control to visualize the heart. Negative control was hybridized with sense zlrrc10 probe (c and f). Arrows indicate the linear heart region that expresses zlrrc10 and cmlc2 . A and V indicate the atrium and ventricle, respectively. (D) Cardiac-specific expression of zlrrc10 in adult zebrafish. RT-PCR analysis was performed using total RNA isolated from three different parts of adult zebrafish as indicated. The zlrrc10 PCR products of 581 bp were visualized by agarose gel electrophoresis. The plasmid pCRII-TOPO- zlrrc10 was used as positive control for the amplification reactions. RT-PCR of β-actin was performed as a loading control.
    Figure Legend Snippet: Cardiac-specific expression of zlrrc10 . (A) The genomic structure of zlrrc10 . The full-length zlrrc10 cDNA consists of two exons. The size of exons, intron and the 5′ and 3′ UTR regions are shown in bp. The amino acid coding regions are represented by the hatched box. (B) An alignment of zebrafish, mouse and human LRRC10 amino acid sequences. Identical residues between all three species are shaded black, and the residues with similar characteristics are shaded gray. The conserved seven LRR motifs are numbered. (C) Cardiac-specific expression of lrrc10 in zebrafish embryos. Zebrafish embryos at 1 dpf (a–c) and 2 dpf (d–f) were subjected to whole-mount in situ hybridization using digoxigenin labeled zlrrc10 antisense cRNA probes (a, d). cmlc2 antisense cRNA probe (b and e) was used for positive control to visualize the heart. Negative control was hybridized with sense zlrrc10 probe (c and f). Arrows indicate the linear heart region that expresses zlrrc10 and cmlc2 . A and V indicate the atrium and ventricle, respectively. (D) Cardiac-specific expression of zlrrc10 in adult zebrafish. RT-PCR analysis was performed using total RNA isolated from three different parts of adult zebrafish as indicated. The zlrrc10 PCR products of 581 bp were visualized by agarose gel electrophoresis. The plasmid pCRII-TOPO- zlrrc10 was used as positive control for the amplification reactions. RT-PCR of β-actin was performed as a loading control.

    Techniques Used: Expressing, In Situ Hybridization, Labeling, Positive Control, Negative Control, Reverse Transcription Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Plasmid Preparation, Amplification

    5) Product Images from "Neisseria meningitidis RTX Proteins Are Not Required for Virulence in Infant Rats "

    Article Title: Neisseria meningitidis RTX Proteins Are Not Required for Virulence in Infant Rats

    Journal: Infection and Immunity

    doi: 10.1128/IAI.71.4.2253-2257.2003

    Scheme of the construction of an MC58 frpC ). The unique Sma ), whereas the kanamycin casette was obtained by amplifying the aphA-3 gene. The subcloning step, for the construction of the frpC mutant, was necessary because the pCRII-TOPO vector contains a kanamycin resistance gene.
    Figure Legend Snippet: Scheme of the construction of an MC58 frpC ). The unique Sma ), whereas the kanamycin casette was obtained by amplifying the aphA-3 gene. The subcloning step, for the construction of the frpC mutant, was necessary because the pCRII-TOPO vector contains a kanamycin resistance gene.

    Techniques Used: Subcloning, Mutagenesis, Plasmid Preparation

    Related Articles

    Clone Assay:

    Article Title: A computational and experimental approach toward a priori identification of alternatively spliced exons
    Article Snippet: .. Each PCR product was excised from the gel, cloned into the pCRII-TOPO vector (Invitrogen), and sequenced. ..

    Article Title: Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target
    Article Snippet: .. RNA Silencing A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR was amplified by PCR (forward primer 5′-CTATTTGGCTAGACGTCTGT-3′ and reverse primer 5′-AATACAGTTTCCTTCCCGTT-3′) and cloned into PCRII-TOPO vector (Invitrogen, http://www.invitrogen.com ). .. The resulting construct was linearized using XhoI (for SP6 RNA polymerase transcription) and SacI (for T7 RNA polymerase transcription).

    Article Title: The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation
    Article Snippet: .. PCR products were cloned into pCRII-TOPO vector with the TOPO TA Cloning kit (K4520-01, Invitrogen/ ThermoFisher Scientific, Waltham, MA, USA) and then sequenced. ..

    Article Title: A new Drosophila POU gene, pdm3, acts in odor receptor expression and axon targeting of olfactory neurons
    Article Snippet: .. The short form was made by using two BsmI (New England Biolabs) sites that flank the alternatively spliced region to swap the long splice form DNA out of the construct and replace it with short splice form DNA which had been cloned previously into the pCRII TOPO vector (Invitrogen) in initial RT-PCR analysis. .. Clones were sequenced and those clones with the correct sequence were swapped into pUAST (Invitrogen) using the restriction enzymes BglII and NotI (New England Biolabs).

    Article Title: Human NMD ensues independently of stable ribosome stalling
    Article Snippet: .. The fused product was then TOPO TA cloned into a pCRII-TOPO vector (Invitrogen) and ligated into the eukaryotic expression vector pcDNA 3.1 (-) using the restriction sites Apa I and Kpn I. ..

    Article Title: Neisseria meningitidis RTX Proteins Are Not Required for Virulence in Infant Rats
    Article Snippet: .. The amplified fragments, upstream and downstream, for frpC and frpC -like, respectively, were ligated and subsequently cloned into the pCRII-TOPO vector (Invitrogen) (Fig. ). .. A kanamycin resistance cassette was inserted between the two fragments amplified for the frpC allele, whereas the Ω fragment encoding resistance to streptomycin was inserted between the two fragments amplified for the frpC -like allele (Fig. ).

    Amplification:

    Article Title: Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target
    Article Snippet: .. RNA Silencing A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR was amplified by PCR (forward primer 5′-CTATTTGGCTAGACGTCTGT-3′ and reverse primer 5′-AATACAGTTTCCTTCCCGTT-3′) and cloned into PCRII-TOPO vector (Invitrogen, http://www.invitrogen.com ). .. The resulting construct was linearized using XhoI (for SP6 RNA polymerase transcription) and SacI (for T7 RNA polymerase transcription).

    Article Title: Use of Leishmania donovani Field Isolates Expressing the Luciferase Reporter Gene in In Vitro Drug Screening †
    Article Snippet: .. The resulting amplicon (1.7 kb) was ligated into the pCRII-TOPO vector (Invitrogen) to create plasmid pCRT- luc and was selected for ampicillin (100 μg/ml) resistance in the presence of 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside and isopropyl-β- d -thiogalactopyranoside. .. Two clones (pCRT- luc 4 and pCRT- luc 8) were sequenced completely in both directions by the dideoxy method to confirm the sequence of the amplicon and to detect the presence of any deletions, additions, or point mutations in the sequence that may have appeared during the amplification of the gene.

    Article Title: Neisseria meningitidis RTX Proteins Are Not Required for Virulence in Infant Rats
    Article Snippet: .. The amplified fragments, upstream and downstream, for frpC and frpC -like, respectively, were ligated and subsequently cloned into the pCRII-TOPO vector (Invitrogen) (Fig. ). .. A kanamycin resistance cassette was inserted between the two fragments amplified for the frpC allele, whereas the Ω fragment encoding resistance to streptomycin was inserted between the two fragments amplified for the frpC -like allele (Fig. ).

    Polymerase Chain Reaction:

    Article Title: A computational and experimental approach toward a priori identification of alternatively spliced exons
    Article Snippet: .. Each PCR product was excised from the gel, cloned into the pCRII-TOPO vector (Invitrogen), and sequenced. ..

    Article Title: Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target
    Article Snippet: .. RNA Silencing A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR was amplified by PCR (forward primer 5′-CTATTTGGCTAGACGTCTGT-3′ and reverse primer 5′-AATACAGTTTCCTTCCCGTT-3′) and cloned into PCRII-TOPO vector (Invitrogen, http://www.invitrogen.com ). .. The resulting construct was linearized using XhoI (for SP6 RNA polymerase transcription) and SacI (for T7 RNA polymerase transcription).

    Article Title: Lrrc10 is required for early heart development and function in zebrafish
    Article Snippet: .. The full-length PCR product of 973 bp consisting of the entire coding sequence, 5′ and 3′ untranslated region was subcloned into pCRII-TOPO vector (Invitrogen) followed by sequencing. .. The nucleotide sequence of the cloned zlrrc10 is identical to the reported sequence.

    Article Title: The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation
    Article Snippet: .. PCR products were cloned into pCRII-TOPO vector with the TOPO TA Cloning kit (K4520-01, Invitrogen/ ThermoFisher Scientific, Waltham, MA, USA) and then sequenced. ..

    TA Cloning:

    Article Title: The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation
    Article Snippet: .. PCR products were cloned into pCRII-TOPO vector with the TOPO TA Cloning kit (K4520-01, Invitrogen/ ThermoFisher Scientific, Waltham, MA, USA) and then sequenced. ..

    Construct:

    Article Title: A new Drosophila POU gene, pdm3, acts in odor receptor expression and axon targeting of olfactory neurons
    Article Snippet: .. The short form was made by using two BsmI (New England Biolabs) sites that flank the alternatively spliced region to swap the long splice form DNA out of the construct and replace it with short splice form DNA which had been cloned previously into the pCRII TOPO vector (Invitrogen) in initial RT-PCR analysis. .. Clones were sequenced and those clones with the correct sequence were swapped into pUAST (Invitrogen) using the restriction enzymes BglII and NotI (New England Biolabs).

    Sequencing:

    Article Title: Lrrc10 is required for early heart development and function in zebrafish
    Article Snippet: .. The full-length PCR product of 973 bp consisting of the entire coding sequence, 5′ and 3′ untranslated region was subcloned into pCRII-TOPO vector (Invitrogen) followed by sequencing. .. The nucleotide sequence of the cloned zlrrc10 is identical to the reported sequence.

    Expressing:

    Article Title: Human NMD ensues independently of stable ribosome stalling
    Article Snippet: .. The fused product was then TOPO TA cloned into a pCRII-TOPO vector (Invitrogen) and ligated into the eukaryotic expression vector pcDNA 3.1 (-) using the restriction sites Apa I and Kpn I. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A new Drosophila POU gene, pdm3, acts in odor receptor expression and axon targeting of olfactory neurons
    Article Snippet: .. The short form was made by using two BsmI (New England Biolabs) sites that flank the alternatively spliced region to swap the long splice form DNA out of the construct and replace it with short splice form DNA which had been cloned previously into the pCRII TOPO vector (Invitrogen) in initial RT-PCR analysis. .. Clones were sequenced and those clones with the correct sequence were swapped into pUAST (Invitrogen) using the restriction enzymes BglII and NotI (New England Biolabs).

    Plasmid Preparation:

    Article Title: A computational and experimental approach toward a priori identification of alternatively spliced exons
    Article Snippet: .. Each PCR product was excised from the gel, cloned into the pCRII-TOPO vector (Invitrogen), and sequenced. ..

    Article Title: Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target
    Article Snippet: .. RNA Silencing A 500-nucleotide fragment (bp 1364–1866) of S. mansoni TGR was amplified by PCR (forward primer 5′-CTATTTGGCTAGACGTCTGT-3′ and reverse primer 5′-AATACAGTTTCCTTCCCGTT-3′) and cloned into PCRII-TOPO vector (Invitrogen, http://www.invitrogen.com ). .. The resulting construct was linearized using XhoI (for SP6 RNA polymerase transcription) and SacI (for T7 RNA polymerase transcription).

    Article Title: Lrrc10 is required for early heart development and function in zebrafish
    Article Snippet: .. The full-length PCR product of 973 bp consisting of the entire coding sequence, 5′ and 3′ untranslated region was subcloned into pCRII-TOPO vector (Invitrogen) followed by sequencing. .. The nucleotide sequence of the cloned zlrrc10 is identical to the reported sequence.

    Article Title: The C/ebp-Atf response element (CARE) location reveals two distinct Atf4-dependent, elongation-mediated mechanisms for transcriptional induction of aminoacyl-tRNA synthetase genes in response to amino acid limitation
    Article Snippet: .. PCR products were cloned into pCRII-TOPO vector with the TOPO TA Cloning kit (K4520-01, Invitrogen/ ThermoFisher Scientific, Waltham, MA, USA) and then sequenced. ..

    Article Title: Use of Leishmania donovani Field Isolates Expressing the Luciferase Reporter Gene in In Vitro Drug Screening †
    Article Snippet: .. The resulting amplicon (1.7 kb) was ligated into the pCRII-TOPO vector (Invitrogen) to create plasmid pCRT- luc and was selected for ampicillin (100 μg/ml) resistance in the presence of 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside and isopropyl-β- d -thiogalactopyranoside. .. Two clones (pCRT- luc 4 and pCRT- luc 8) were sequenced completely in both directions by the dideoxy method to confirm the sequence of the amplicon and to detect the presence of any deletions, additions, or point mutations in the sequence that may have appeared during the amplification of the gene.

    Article Title: A new Drosophila POU gene, pdm3, acts in odor receptor expression and axon targeting of olfactory neurons
    Article Snippet: .. The short form was made by using two BsmI (New England Biolabs) sites that flank the alternatively spliced region to swap the long splice form DNA out of the construct and replace it with short splice form DNA which had been cloned previously into the pCRII TOPO vector (Invitrogen) in initial RT-PCR analysis. .. Clones were sequenced and those clones with the correct sequence were swapped into pUAST (Invitrogen) using the restriction enzymes BglII and NotI (New England Biolabs).

    Article Title: Human NMD ensues independently of stable ribosome stalling
    Article Snippet: .. The fused product was then TOPO TA cloned into a pCRII-TOPO vector (Invitrogen) and ligated into the eukaryotic expression vector pcDNA 3.1 (-) using the restriction sites Apa I and Kpn I. ..

    Article Title: Neisseria meningitidis RTX Proteins Are Not Required for Virulence in Infant Rats
    Article Snippet: .. The amplified fragments, upstream and downstream, for frpC and frpC -like, respectively, were ligated and subsequently cloned into the pCRII-TOPO vector (Invitrogen) (Fig. ). .. A kanamycin resistance cassette was inserted between the two fragments amplified for the frpC allele, whereas the Ω fragment encoding resistance to streptomycin was inserted between the two fragments amplified for the frpC -like allele (Fig. ).

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    Thermo Fisher pcrii topo vector
    Pcrii Topo Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcrii topo vector/product/Thermo Fisher
    Average 94 stars, based on 800 article reviews
    Price from $9.99 to $1999.99
    pcrii topo vector - by Bioz Stars, 2020-08
    94/100 stars
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