mrna molecules  (Thermo Fisher)


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    Structured Review

    Thermo Fisher mrna molecules
    Exogenous <t>Oct4</t> expression shows robust nuclear localization. (A) Experimental scheme for the production of OCT4 overexpressing and depleted embryos. (B) IF analysis of embryos injected with Oct4 <t>mRNA</t> of various concentrations at 6 h after injection. Representative images of maximum projection and intensity plots in OCT4-OE and control embryos were shown. The same strong laser intensity was applied to all samples. Dotted and solid lines indicate DAPI and OCT4 signals respectively. The scale bars represent 20 μm. (C) SDS-WB analysis using cytoplasmic and nuclear lysates from zygotes of OCT4-OE and control samples. Two independent experiments were conducted. RNAPII was used as a loading control. (D) Quantification of OCT4 nuclear protein levels. The same laser intensity was applied to all samples, and the signal intensities were measured using ImageJ. Representative images are presented, and quantified signals are shown as bar graph. n indicates the number of cells analyzed for each measurement. The P -values were calculated by Student’s t -test. Error bars indicate the standard deviation. Scale bars = 20 μm.
    Mrna Molecules, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mrna molecules - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "Spatiotemporal dynamics of OCT4 protein localization during preimplantation development in mice"

    Article Title: Spatiotemporal dynamics of OCT4 protein localization during preimplantation development in mice

    Journal: Reproduction (Cambridge, England)

    doi: 10.1530/REP-16-0277

    Exogenous Oct4 expression shows robust nuclear localization. (A) Experimental scheme for the production of OCT4 overexpressing and depleted embryos. (B) IF analysis of embryos injected with Oct4 mRNA of various concentrations at 6 h after injection. Representative images of maximum projection and intensity plots in OCT4-OE and control embryos were shown. The same strong laser intensity was applied to all samples. Dotted and solid lines indicate DAPI and OCT4 signals respectively. The scale bars represent 20 μm. (C) SDS-WB analysis using cytoplasmic and nuclear lysates from zygotes of OCT4-OE and control samples. Two independent experiments were conducted. RNAPII was used as a loading control. (D) Quantification of OCT4 nuclear protein levels. The same laser intensity was applied to all samples, and the signal intensities were measured using ImageJ. Representative images are presented, and quantified signals are shown as bar graph. n indicates the number of cells analyzed for each measurement. The P -values were calculated by Student’s t -test. Error bars indicate the standard deviation. Scale bars = 20 μm.
    Figure Legend Snippet: Exogenous Oct4 expression shows robust nuclear localization. (A) Experimental scheme for the production of OCT4 overexpressing and depleted embryos. (B) IF analysis of embryos injected with Oct4 mRNA of various concentrations at 6 h after injection. Representative images of maximum projection and intensity plots in OCT4-OE and control embryos were shown. The same strong laser intensity was applied to all samples. Dotted and solid lines indicate DAPI and OCT4 signals respectively. The scale bars represent 20 μm. (C) SDS-WB analysis using cytoplasmic and nuclear lysates from zygotes of OCT4-OE and control samples. Two independent experiments were conducted. RNAPII was used as a loading control. (D) Quantification of OCT4 nuclear protein levels. The same laser intensity was applied to all samples, and the signal intensities were measured using ImageJ. Representative images are presented, and quantified signals are shown as bar graph. n indicates the number of cells analyzed for each measurement. The P -values were calculated by Student’s t -test. Error bars indicate the standard deviation. Scale bars = 20 μm.

    Techniques Used: Expressing, Injection, Western Blot, Standard Deviation

    In-depth expression analysis of Oct4 mRNA and protein in oocytes and preimplantation embryos. (A) RT-PCR analysis from the transcription start site (TSS) to translation termination site in oocytes and preimplantation embryos. Two independent experiments were conducted. (B) Single-cell qPCR analysis for two Oct4 exon junction regions. n, the number of cells used for the analysis. * P
    Figure Legend Snippet: In-depth expression analysis of Oct4 mRNA and protein in oocytes and preimplantation embryos. (A) RT-PCR analysis from the transcription start site (TSS) to translation termination site in oocytes and preimplantation embryos. Two independent experiments were conducted. (B) Single-cell qPCR analysis for two Oct4 exon junction regions. n, the number of cells used for the analysis. * P

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    2) Product Images from "A Conditional Mouse Mutant in the Tumor Suppressor SdhD Gene Unveils a Link between p21WAF1/Cip1 Induction and Mitochondrial Dysfunction"

    Article Title: A Conditional Mouse Mutant in the Tumor Suppressor SdhD Gene Unveils a Link between p21WAF1/Cip1 Induction and Mitochondrial Dysfunction

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0085528

    Functional analysis and validation of the microarray. A. Comparative functional analysis of datasets generated from adrenal medulla (AM) and kidney samples from SDHD-ESR mice. The significance of each molecular and cellular function is indicated by –log of the p-value. B. Quantitative RT-PCR of RNA samples used in the microarray study based on specific primers for amplification of the Cdkn1a mRNA.
    Figure Legend Snippet: Functional analysis and validation of the microarray. A. Comparative functional analysis of datasets generated from adrenal medulla (AM) and kidney samples from SDHD-ESR mice. The significance of each molecular and cellular function is indicated by –log of the p-value. B. Quantitative RT-PCR of RNA samples used in the microarray study based on specific primers for amplification of the Cdkn1a mRNA.

    Techniques Used: Functional Assay, Microarray, Generated, Electron Paramagnetic Resonance, Mouse Assay, Cell Function Assay, Quantitative RT-PCR, Amplification

    3) Product Images from "Novel insights into RNP granules by employing the trypanosome's microtubule skeleton as a molecular sieve"

    Article Title: Novel insights into RNP granules by employing the trypanosome's microtubule skeleton as a molecular sieve

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkv731

    mRNA FISH using the Affymetrix system. ( A ) Single mRNA FISH using Affymetrix probe sets antisense to RPL7a (left) or DBP1 (right) together with an oligo antisense to the mini-exon (ME). One representative untreated and starved cell out of several experiments is shown for each oligo set as Z-stack projections. Further images are shown in Supplementary Figure S6. As a negative control, we used an Affymetrix probe set sense to RPL7a and an oligo sense to the mini-exon sequence, both probes gave no or very weak signals (data not shown). ( B ) The number of RPL7a and DBP1 mRNA molecules (spots) per starved cell that showed no or total/partial colocalisation with a starvation stress granule is shown as bar chart. Data of two independent experiments are shown.
    Figure Legend Snippet: mRNA FISH using the Affymetrix system. ( A ) Single mRNA FISH using Affymetrix probe sets antisense to RPL7a (left) or DBP1 (right) together with an oligo antisense to the mini-exon (ME). One representative untreated and starved cell out of several experiments is shown for each oligo set as Z-stack projections. Further images are shown in Supplementary Figure S6. As a negative control, we used an Affymetrix probe set sense to RPL7a and an oligo sense to the mini-exon sequence, both probes gave no or very weak signals (data not shown). ( B ) The number of RPL7a and DBP1 mRNA molecules (spots) per starved cell that showed no or total/partial colocalisation with a starvation stress granule is shown as bar chart. Data of two independent experiments are shown.

    Techniques Used: Fluorescence In Situ Hybridization, Negative Control, Sequencing

    4) Product Images from "Inhibition of Xenograft Tumor Growth by Gold Nanoparticle-DNA Oligonucleotide Conjugates-Assisted Delivery of BAX mRNA"

    Article Title: Inhibition of Xenograft Tumor Growth by Gold Nanoparticle-DNA Oligonucleotide Conjugates-Assisted Delivery of BAX mRNA

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0075369

    Influence of BAX mRNA delivery via AuNP-DNA conjugate or liposome on cell viability and mRNA half-life. (A) Relative viability of 5′BAX mRNA-transfected cells. HeLa cells were incubated with AuNP-αRNA I-5’BAX-null mRNA (Bax-null mRNA), AuNP-αRNA I-5’BAX mRNA, or liposome for 24 h. (B) RT-PCR analysis of 5′BAX mRNA to measure mRNA half-life. Amplification of cDNA from 5′BAX mRNA was achieved using a PCR primer (RNA I-AUG), designed specifically to bind to the 5’ UTR of 5′BAX mRNA, which is complementary to the cargo DNA of the AuNP-DNA. Equal loading was ensured using an internal control (GAPDH). The values were normalized against control cell values and are shown as the mean ± SEM (standard error of mean). Experiments were performed in triplicate and repeated at least three times.
    Figure Legend Snippet: Influence of BAX mRNA delivery via AuNP-DNA conjugate or liposome on cell viability and mRNA half-life. (A) Relative viability of 5′BAX mRNA-transfected cells. HeLa cells were incubated with AuNP-αRNA I-5’BAX-null mRNA (Bax-null mRNA), AuNP-αRNA I-5’BAX mRNA, or liposome for 24 h. (B) RT-PCR analysis of 5′BAX mRNA to measure mRNA half-life. Amplification of cDNA from 5′BAX mRNA was achieved using a PCR primer (RNA I-AUG), designed specifically to bind to the 5’ UTR of 5′BAX mRNA, which is complementary to the cargo DNA of the AuNP-DNA. Equal loading was ensured using an internal control (GAPDH). The values were normalized against control cell values and are shown as the mean ± SEM (standard error of mean). Experiments were performed in triplicate and repeated at least three times.

    Techniques Used: Transfection, Incubation, Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction

    Delivery of dsRED mRNA to HeLa cells by AuNP-DNA conjugates. (A) Measurement of the loading capacity of AuNP-αRNA I for 5′dsRED mRNA and 5′∆RNA I-dsRED mRNA. Known amounts of 5’ dsRED mRNA or 5′∆RNA I-dsRED mRNA (5, 10, and 20 pmol) and ten µl of varying concentrations (2, 4, and 8 µM) of these mRNAs hybridized to AuNP-DNA were loaded (lanes 4-6) to assess the loading capacity of for 5’ dsRED mRNA containing a sequence complementary to αRNA I at the 5′-terminus. The concentration of AuNP-DNA was 10 nM. Amounts of mRNA bound to AuNP-DNA at each concentration were plotted and shown in the graph. (B) Efficient delivery of AuNP-αRNA I-Cy3-5 ′dsRED mRNA into HeLa cells. Cells were treated with AuNP-αRNA I alone, AuNP-αRNA I-Cy3-5’∆RNA I-dsRED mRNA, or AuNP-αRNA I-Cy3-5’dsRED mRNA; these are represented schematically in diagrams. The final concentrations of AuNP-αRNA I and dsRED mRNA were 1 nM and 0.2 µM, respectively. Representative confocal fluorescence microscopy images of HeLa cells detecting Cy3-5’dsRED and Cy3-5’∆RNA I-dsRED mRNA (a) and dsRED protein (b) are shown. Transmission images of cells (c) and overlay images of a, b, and c (d) are also shown. Bar indicates 20 µm.
    Figure Legend Snippet: Delivery of dsRED mRNA to HeLa cells by AuNP-DNA conjugates. (A) Measurement of the loading capacity of AuNP-αRNA I for 5′dsRED mRNA and 5′∆RNA I-dsRED mRNA. Known amounts of 5’ dsRED mRNA or 5′∆RNA I-dsRED mRNA (5, 10, and 20 pmol) and ten µl of varying concentrations (2, 4, and 8 µM) of these mRNAs hybridized to AuNP-DNA were loaded (lanes 4-6) to assess the loading capacity of for 5’ dsRED mRNA containing a sequence complementary to αRNA I at the 5′-terminus. The concentration of AuNP-DNA was 10 nM. Amounts of mRNA bound to AuNP-DNA at each concentration were plotted and shown in the graph. (B) Efficient delivery of AuNP-αRNA I-Cy3-5 ′dsRED mRNA into HeLa cells. Cells were treated with AuNP-αRNA I alone, AuNP-αRNA I-Cy3-5’∆RNA I-dsRED mRNA, or AuNP-αRNA I-Cy3-5’dsRED mRNA; these are represented schematically in diagrams. The final concentrations of AuNP-αRNA I and dsRED mRNA were 1 nM and 0.2 µM, respectively. Representative confocal fluorescence microscopy images of HeLa cells detecting Cy3-5’dsRED and Cy3-5’∆RNA I-dsRED mRNA (a) and dsRED protein (b) are shown. Transmission images of cells (c) and overlay images of a, b, and c (d) are also shown. Bar indicates 20 µm.

    Techniques Used: Sequencing, Concentration Assay, Fluorescence, Microscopy, Transmission Assay

    Effects of directionality of mRNA binding to AuNP-DNA and presence of cap structure at the 5′-terminus of mRNA. HeLa cells were treated with AuNP-αRNA I hybridized to Cy3-5’dsRED mRNA, Cy3-3’dsRED mRNA, Cy3-5′, 3’dsRED mRNA, or Cy3-CAP-5’dsRED mRNA. Fluorescent images show the Cy3-labeled mRNA signal in the left panels and the red fluorescent signal of the dsRED protein in the right panels. Representative confocal microscopy images of HeLa cells for each treatment are shown, along with schematic representations of in vitro transcripts and a diagram of the AuNP-αRNA I-mRNA binding structure. Bar indicates 20 µm. Relative intensities of fluorescence in cells were measured using ImageJ software. The values are shown by setting intensities of fluorescence from cells treated with AuNP-αRNA I + Cy3-5’dsRED mRNA as 100. Relative intensities of fluorescence were obtained from 10 cells of each treatment and are shown as the mean ± SEM (standard error of mean).
    Figure Legend Snippet: Effects of directionality of mRNA binding to AuNP-DNA and presence of cap structure at the 5′-terminus of mRNA. HeLa cells were treated with AuNP-αRNA I hybridized to Cy3-5’dsRED mRNA, Cy3-3’dsRED mRNA, Cy3-5′, 3’dsRED mRNA, or Cy3-CAP-5’dsRED mRNA. Fluorescent images show the Cy3-labeled mRNA signal in the left panels and the red fluorescent signal of the dsRED protein in the right panels. Representative confocal microscopy images of HeLa cells for each treatment are shown, along with schematic representations of in vitro transcripts and a diagram of the AuNP-αRNA I-mRNA binding structure. Bar indicates 20 µm. Relative intensities of fluorescence in cells were measured using ImageJ software. The values are shown by setting intensities of fluorescence from cells treated with AuNP-αRNA I + Cy3-5’dsRED mRNA as 100. Relative intensities of fluorescence were obtained from 10 cells of each treatment and are shown as the mean ± SEM (standard error of mean).

    Techniques Used: Binding Assay, Labeling, Confocal Microscopy, In Vitro, Fluorescence, Software

    5) Product Images from "A-to-I RNA editing in the rat brain is age-dependent, region-specific and sensitive to environmental stress across generations"

    Article Title: A-to-I RNA editing in the rat brain is age-dependent, region-specific and sensitive to environmental stress across generations

    Journal: BMC Genomics

    doi: 10.1186/s12864-017-4409-8

    Transgenerational effects of stress on A-to-I RNA editing and related gene expression levels in the rat brain. a Experimental Design: Transgenerational transmission of stress effects. b mRNA gene expression of Adar , Adarb1 and Htr2c ( b 1 ), and Htr2c A-D site editing ( b 2 ) In PFC of F1 (top row) and F2 (bottom row) offspring. c mRNA gene expression of Adar , Adarb1 and Htr2c ( c 1 ), and RNA editing levels of 4 Htr2c sites ( c 2 ), In AMY of F1 (top row) and F2 (bottom row) offspring. d Htr2c isoform distribution changes between C and PRS (F0), O1-C and O1-PRS (F1) and O2-C and O2-PRS (F2) in PFC ( d 1 ) and AMY ( d 2 ). * p
    Figure Legend Snippet: Transgenerational effects of stress on A-to-I RNA editing and related gene expression levels in the rat brain. a Experimental Design: Transgenerational transmission of stress effects. b mRNA gene expression of Adar , Adarb1 and Htr2c ( b 1 ), and Htr2c A-D site editing ( b 2 ) In PFC of F1 (top row) and F2 (bottom row) offspring. c mRNA gene expression of Adar , Adarb1 and Htr2c ( c 1 ), and RNA editing levels of 4 Htr2c sites ( c 2 ), In AMY of F1 (top row) and F2 (bottom row) offspring. d Htr2c isoform distribution changes between C and PRS (F0), O1-C and O1-PRS (F1) and O2-C and O2-PRS (F2) in PFC ( d 1 ) and AMY ( d 2 ). * p

    Techniques Used: Expressing, Transmission Assay

    Age-dependent changes in A-to-I RNA editing and gene expression in rat brain. a Significant age-dependent changes in RNA editing at non-synonymous editing sites in PFC ( a 1 ) and AMY ( a 2 ). b Age-dependent changes in Adar and Adarb1 mRNA expression in PFC ( b 1 ) and AMY ( b 2 ). c Age-dependent changes in Htr2c A-D site editing ( c 1 ) and mRNA expression ( c 2 ) in PFC. d Age-dependent changes in Htr2c A-D site editing ( d 1 ) and mRNA expression ( d 2 ) in AMY. e Age-dependent changes in Htr2c isoform prevalence in PFC ( e 1 ) and AMY ( e 2 ). Columns represent individual samples. Rows represent editing sites. * p
    Figure Legend Snippet: Age-dependent changes in A-to-I RNA editing and gene expression in rat brain. a Significant age-dependent changes in RNA editing at non-synonymous editing sites in PFC ( a 1 ) and AMY ( a 2 ). b Age-dependent changes in Adar and Adarb1 mRNA expression in PFC ( b 1 ) and AMY ( b 2 ). c Age-dependent changes in Htr2c A-D site editing ( c 1 ) and mRNA expression ( c 2 ) in PFC. d Age-dependent changes in Htr2c A-D site editing ( d 1 ) and mRNA expression ( d 2 ) in AMY. e Age-dependent changes in Htr2c isoform prevalence in PFC ( e 1 ) and AMY ( e 2 ). Columns represent individual samples. Rows represent editing sites. * p

    Techniques Used: Expressing

    Stress-induced changes in A-to-I RNA editing and related gene expression levels in rat brain. a stress-induced changes in Adar and Adarb1 mRNA expression ( a 1 ), RNA editing ( a 2 ) and Htr2c mRNA expression ( a 3 ) in PFC. b stress-induced changes in Adar and Adarb1 mRNA expression ( b 1 ), RNA editing ( b 2 ) and Htr2c mRNA expression ( b 3 ) in AMY. Only significant editing changes are shown. * p
    Figure Legend Snippet: Stress-induced changes in A-to-I RNA editing and related gene expression levels in rat brain. a stress-induced changes in Adar and Adarb1 mRNA expression ( a 1 ), RNA editing ( a 2 ) and Htr2c mRNA expression ( a 3 ) in PFC. b stress-induced changes in Adar and Adarb1 mRNA expression ( b 1 ), RNA editing ( b 2 ) and Htr2c mRNA expression ( b 3 ) in AMY. Only significant editing changes are shown. * p

    Techniques Used: Expressing

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: A Conditional Mouse Mutant in the Tumor Suppressor SdhD Gene Unveils a Link between p21WAF1/Cip1 Induction and Mitochondrial Dysfunction
    Article Snippet: .. Reverse transcription of mRNA was performed with the Superscript II reverse transcriptase kit (Life Technologies), and specific mRNA molecules were amplified by quantitative PCR in the presence of SYBR green® (Life Technologies) with the following primers for each gene: SdhD , 5′-CCAGCACATTCACCTGTCA-3′ and 5′-ATCAGCCCCAAGAGCAGAA-3′ ; Vegf , 5′-CGCAAGAAATCCCGGTTTAA-3′ and 5′-CAAATGCTTTCTCCGCTCTGA-3′ ; Glut1 , 5′-CCAGCTGGGAATCGTCGTT-3′ and 5′-CAAGTCTGCATTGCCCATGAT-3′ ; Phd3 , 5′-CAGACCGCAGGAATCCACAT-3′ and 5′-CATCGAAGTACCAGACAGTCATAGC-3′ ; Cdkn1a , 5′-TCCACAGCGATATCCAGACATT-3′ and 5′-CGGACATCACCAGGATTGG-3′ . ..

    Negative Control:

    Article Title: Spatiotemporal dynamics of OCT4 protein localization during preimplantation development in mice
    Article Snippet: .. The same number of mRNA molecules was found in 200 ng/µL Oct4 mRNA and 130 ng/µL EGFP mRNA used for injection. siRNAs targeting Oct4 (sense: 5ʹ-GUU CGA GUA UGG UUC UGU ATT-3ʹ, antisense: 5ʹ-UAC AGA ACC AUA CUC GAA CCA-3ʹ) and a negative control (silencer select negative control, #4390846; Ambion) were purchased from Life Technologies. .. Each siRNA (25 ng/mL) was injected into zygotes.

    Amplification:

    Article Title: A Conditional Mouse Mutant in the Tumor Suppressor SdhD Gene Unveils a Link between p21WAF1/Cip1 Induction and Mitochondrial Dysfunction
    Article Snippet: .. Reverse transcription of mRNA was performed with the Superscript II reverse transcriptase kit (Life Technologies), and specific mRNA molecules were amplified by quantitative PCR in the presence of SYBR green® (Life Technologies) with the following primers for each gene: SdhD , 5′-CCAGCACATTCACCTGTCA-3′ and 5′-ATCAGCCCCAAGAGCAGAA-3′ ; Vegf , 5′-CGCAAGAAATCCCGGTTTAA-3′ and 5′-CAAATGCTTTCTCCGCTCTGA-3′ ; Glut1 , 5′-CCAGCTGGGAATCGTCGTT-3′ and 5′-CAAGTCTGCATTGCCCATGAT-3′ ; Phd3 , 5′-CAGACCGCAGGAATCCACAT-3′ and 5′-CATCGAAGTACCAGACAGTCATAGC-3′ ; Cdkn1a , 5′-TCCACAGCGATATCCAGACATT-3′ and 5′-CGGACATCACCAGGATTGG-3′ . ..

    Article Title: MicroRNA-155 contributes to enhanced resistance to apoptosis in monocytes from patients with rheumatoid arthritis
    Article Snippet: .. For amplification of mRNA molecules, TriZol extracted RNA was reverse transcribed using a High Capacity cDNA Reverse Transcription Kit (Life Technologies) and amplified using SensiFAST SYBR Green Master mix (Bioline, London, UK) on an ABI7900HT machine. .. 2.6 Transfection Transfection of CD14+ cells was performed using the NTER nanoparticle transfection reagent (Sigma, St Louis, MO, USA) following manufacturer's instructions.

    In Vitro:

    Article Title: Inhibition of Xenograft Tumor Growth by Gold Nanoparticle-DNA Oligonucleotide Conjugates-Assisted Delivery of BAX mRNA
    Article Snippet: .. In vitro synthesis of mRNA The mRNA molecules were synthesized from DNA templates containing a T7 promoter followed by a sequence of dsRED , GFP , and BAX using the MEGAscriptTM kit (Ambion, USA), according to the manufacturer’s instructions. .. In vitro synthesized mRNA molecules were comprised of 5′-end Kozak sequences ( 5′-GCCGCCACC-3′ ), an open reading frame (ORF) beginning with a start codon and ending with a stop codon, and a polyA tail consisting of 20 nucleotides (ntds) at the 3′-end.

    Magnetic Beads:

    Article Title: Differential responsiveness to BRAF inhibitors of melanoma cell lines BRAF V600E-mutated
    Article Snippet: .. Enrichment in mRNA molecules was obtained by using oligo (dT) magnetic beads (Ambion® Poly (A) Purist™ MAG Kit). .. After the mRNA was fragmented in short fragments (approximately 200 bp), cDNA was synthesized by random hexamer primers (Illumina TruSeq Stranded mRNA Library Prep Kit).

    Synthesized:

    Article Title: Inhibition of Xenograft Tumor Growth by Gold Nanoparticle-DNA Oligonucleotide Conjugates-Assisted Delivery of BAX mRNA
    Article Snippet: .. In vitro synthesis of mRNA The mRNA molecules were synthesized from DNA templates containing a T7 promoter followed by a sequence of dsRED , GFP , and BAX using the MEGAscriptTM kit (Ambion, USA), according to the manufacturer’s instructions. .. In vitro synthesized mRNA molecules were comprised of 5′-end Kozak sequences ( 5′-GCCGCCACC-3′ ), an open reading frame (ORF) beginning with a start codon and ending with a stop codon, and a polyA tail consisting of 20 nucleotides (ntds) at the 3′-end.

    Article Title: Mapping of Chikungunya Virus Interactions with Host Proteins Identified nsP2 as a Highly Connected Viral Component
    Article Snippet: .. Then, both capped and polyadenylated mRNA molecules encoding firefly luciferase were synthesized from this plasmid using the T7 mMessage mMachine T7 Ultra kit from Ambion following the manufacturer's recommendations. .. Synthesized mRNA molecules were purified with the Qiagen RNeasy minikit.

    SYBR Green Assay:

    Article Title: A Conditional Mouse Mutant in the Tumor Suppressor SdhD Gene Unveils a Link between p21WAF1/Cip1 Induction and Mitochondrial Dysfunction
    Article Snippet: .. Reverse transcription of mRNA was performed with the Superscript II reverse transcriptase kit (Life Technologies), and specific mRNA molecules were amplified by quantitative PCR in the presence of SYBR green® (Life Technologies) with the following primers for each gene: SdhD , 5′-CCAGCACATTCACCTGTCA-3′ and 5′-ATCAGCCCCAAGAGCAGAA-3′ ; Vegf , 5′-CGCAAGAAATCCCGGTTTAA-3′ and 5′-CAAATGCTTTCTCCGCTCTGA-3′ ; Glut1 , 5′-CCAGCTGGGAATCGTCGTT-3′ and 5′-CAAGTCTGCATTGCCCATGAT-3′ ; Phd3 , 5′-CAGACCGCAGGAATCCACAT-3′ and 5′-CATCGAAGTACCAGACAGTCATAGC-3′ ; Cdkn1a , 5′-TCCACAGCGATATCCAGACATT-3′ and 5′-CGGACATCACCAGGATTGG-3′ . ..

    Article Title: MicroRNA-155 contributes to enhanced resistance to apoptosis in monocytes from patients with rheumatoid arthritis
    Article Snippet: .. For amplification of mRNA molecules, TriZol extracted RNA was reverse transcribed using a High Capacity cDNA Reverse Transcription Kit (Life Technologies) and amplified using SensiFAST SYBR Green Master mix (Bioline, London, UK) on an ABI7900HT machine. .. 2.6 Transfection Transfection of CD14+ cells was performed using the NTER nanoparticle transfection reagent (Sigma, St Louis, MO, USA) following manufacturer's instructions.

    Luciferase:

    Article Title: Mapping of Chikungunya Virus Interactions with Host Proteins Identified nsP2 as a Highly Connected Viral Component
    Article Snippet: .. Then, both capped and polyadenylated mRNA molecules encoding firefly luciferase were synthesized from this plasmid using the T7 mMessage mMachine T7 Ultra kit from Ambion following the manufacturer's recommendations. .. Synthesized mRNA molecules were purified with the Qiagen RNeasy minikit.

    Sequencing:

    Article Title: Inhibition of Xenograft Tumor Growth by Gold Nanoparticle-DNA Oligonucleotide Conjugates-Assisted Delivery of BAX mRNA
    Article Snippet: .. In vitro synthesis of mRNA The mRNA molecules were synthesized from DNA templates containing a T7 promoter followed by a sequence of dsRED , GFP , and BAX using the MEGAscriptTM kit (Ambion, USA), according to the manufacturer’s instructions. .. In vitro synthesized mRNA molecules were comprised of 5′-end Kozak sequences ( 5′-GCCGCCACC-3′ ), an open reading frame (ORF) beginning with a start codon and ending with a stop codon, and a polyA tail consisting of 20 nucleotides (ntds) at the 3′-end.

    Injection:

    Article Title: Spatiotemporal dynamics of OCT4 protein localization during preimplantation development in mice
    Article Snippet: .. The same number of mRNA molecules was found in 200 ng/µL Oct4 mRNA and 130 ng/µL EGFP mRNA used for injection. siRNAs targeting Oct4 (sense: 5ʹ-GUU CGA GUA UGG UUC UGU ATT-3ʹ, antisense: 5ʹ-UAC AGA ACC AUA CUC GAA CCA-3ʹ) and a negative control (silencer select negative control, #4390846; Ambion) were purchased from Life Technologies. .. Each siRNA (25 ng/mL) was injected into zygotes.

    Plasmid Preparation:

    Article Title: Mapping of Chikungunya Virus Interactions with Host Proteins Identified nsP2 as a Highly Connected Viral Component
    Article Snippet: .. Then, both capped and polyadenylated mRNA molecules encoding firefly luciferase were synthesized from this plasmid using the T7 mMessage mMachine T7 Ultra kit from Ambion following the manufacturer's recommendations. .. Synthesized mRNA molecules were purified with the Qiagen RNeasy minikit.

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    Thermo Fisher mrna molecules
    Mrna Molecules, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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