reagents rna isolation Search Results


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  • 99
    Thermo Fisher trizol rna isolation reagents
    Total <t>RNA</t> was extracted from the aorta of different groups using <t>TRIzol</t> reagent; MiR-1 expression was determined using the miRNA plate assay kit; for normalized RNA content, the U6 snRNA was the internal control. a miR-1 expression was detected in control and AS mice. b miR-1 treatment influences miR-1 expression in the aorta. Levels of miR-1 were detected in different miR-treated AS mice, * P
    Trizol Rna Isolation Reagents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trizol rna isolation reagents/product/Thermo Fisher
    Average 99 stars, based on 423 article reviews
    Price from $9.99 to $1999.99
    trizol rna isolation reagents - by Bioz Stars, 2019-10
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    99
    Qiagen total rna isolation reagent paxgene blood rna kit
    Total <t>RNA</t> was extracted from the aorta of different groups using <t>TRIzol</t> reagent; MiR-1 expression was determined using the miRNA plate assay kit; for normalized RNA content, the U6 snRNA was the internal control. a miR-1 expression was detected in control and AS mice. b miR-1 treatment influences miR-1 expression in the aorta. Levels of miR-1 were detected in different miR-treated AS mice, * P
    Total Rna Isolation Reagent Paxgene Blood Rna Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/total rna isolation reagent paxgene blood rna kit/product/Qiagen
    Average 99 stars, based on 6 article reviews
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    99
    Thermo Fisher plant rna isolation reagent
    Custom microarray analysis of the accumulation of antisense <t>RNAs</t> in wild-type and rdr1/2/6 plants subjected to drought stress. A, MA plots illustrating the accumulation level of poly (A-) sense <t>RNA</t> and poly (A-) antisense RNA at 7,138 gene loci in which
    Plant Rna Isolation Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plant rna isolation reagent/product/Thermo Fisher
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    93
    Tel Test Inc rna bee rna isolation reagent
    Temporal expression of a poCI mRNA. <t>RT-PCR</t> of <t>RNA</t> isolated from whole embryos between stage 4 and stage 32. Chordin serves as a staging control and Ornithine decarboxylase (ODC) as a control for input RNA levels.
    Rna Bee Rna Isolation Reagent, supplied by Tel Test Inc, used in various techniques. Bioz Stars score: 93/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna bee rna isolation reagent/product/Tel Test Inc
    Average 93 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
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    92
    Roche tripure rna isolation reagent
    Co-culture of hMSCs transduced with ADAMTS5-targeting shRNA effectively reduces ADAMTS5 expression in flow cytometry sorted RFP-labeled SW982. (A) Following co-culture of non-targeting control shRNA or ADAMTS5-targeting shRNA transduced hMSCs with RFP labeled SW982 cells, the cells were sorted by flow cytometry. The gating threshold (RFP+ sorted) was set based on the analysis of co-cultured hMSCs with unlabeled SW982 cells (no SW982-RFP cells). RFP-positive SW982 cells that were co-cultured with hMSCs previously transduced with non-targeting control-shRNA or ADAMTS5-targeting shRNA were sorted directly into <t>TriPure</t> reagent for <t>RNA</t> isolation. (B) The histogram shows the relative gene expression for ADAMTS5 in the RFP-positive cell population isolated after co-culture with hMSC-shRNA-control or hMSC-shRNA-ADAMTS5 cells as determined by quantitative real time RT-PCR. The result is from a representative experiment.
    Tripure Rna Isolation Reagent, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tripure rna isolation reagent/product/Roche
    Average 92 stars, based on 93 article reviews
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    tripure rna isolation reagent - by Bioz Stars, 2019-10
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    83
    tiangen biotech co plant rna isolation reagent
    Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR. A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total <t>RNA</t> was isolated respectively from A. <t>annua</t> leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1 - RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
    Plant Rna Isolation Reagent, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 83/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 83 stars, based on 12 article reviews
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    83
    Biotex Laboratories total rna isolation reagent
    MARCH2 expression is required for carvedilol-induced ubiquitination, lysosomal trafficking, and degradation of endogenous β 2 ARs. (A) Rat VSMCs were transfected with either control (CTL) or MARCH2 <t>siRNA;</t> total <t>RNA</t> was extracted 48 h later and subjected to RT-PCR for MARCH2 or GAPDH, as indicated. There was an 80 ± 5% knockdown of MARCH2 mRNA ( n = 6). (B) Rat VSMCs transfected with CTL or MARCH2 siRNA were stimulated with carvedilol (1 h, 37°C), and β 2 AR immunoprecipitates were probed with anti-Ub (top) or anti-β 2 AR (bottom) IgG. (C) The ubiquitin signals were quantified, normalized to the β 2 ARs in each IP sample, and plotted as bar graphs (mean ± SEM). Compared with NS: *, P
    Total Rna Isolation Reagent, supplied by Biotex Laboratories, used in various techniques. Bioz Stars score: 83/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 83 stars, based on 3 article reviews
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    total rna isolation reagent - by Bioz Stars, 2019-10
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    83
    AMS Biotechnology rna bee rna isolation reagent
    Northern blot analysis examining gene expression in cells infected with wild-type (WT) or STOP-mutant EHV-1. <t>RK13</t> cells were infected with wild-type EHV-1 or the UL4aa18stop EHV-1, and <t>RNA</t> was isolated at 4, 8, and 12 hours post-infection. Northern blot
    Rna Bee Rna Isolation Reagent, supplied by AMS Biotechnology, used in various techniques. Bioz Stars score: 83/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna bee rna isolation reagent/product/AMS Biotechnology
    Average 83 stars, based on 36 article reviews
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    79
    TEL-TEST Inc total rna mrna isolation reagent rna stat 60
    Small interfering <t>RNA</t> specific for PIP5KI β (but not α or γ ) reduces PIP2 levels in HeLa cells. HeLa cells were transfected with siRNA oligos targeting the α, β, or γ isoforms of PIP5KI or an irrelevant sequence (control, C). Western blots with antibodies specific for the respective protein and tubulin (loading control) were performed 48 h after transfection. (A) An example of a Western blot specific for PIP5KIα is shown. Similar blots were quantified by densitometry, and the values are presented graphically. (B) PIP2 levels in cultures transfected with siRNAs specific for the enzymes shown were measured as described in Fig. 1 B. (C) <t>mRNA</t> for each enzyme in cells treated with siRNA specific for one isoform were measured by real-time PCR. For each isoform, values in A and B are expressed as a percentage of the value measured in the control transfected with unrelated siRNA. All graphs present the averages of at least three experiments with SD.
    Total Rna Mrna Isolation Reagent Rna Stat 60, supplied by TEL-TEST Inc, used in various techniques. Bioz Stars score: 79/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 79 stars, based on 17 article reviews
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    79
    Bioteke Corporation plant rna isolation reagent
    Cloning, expression and purification of AAL-2 ( A ) Total <t>RNA</t> was isolated from fresh A. <t>aegerita</t> fruiting body and amplified by reverse transcription–PCR. The AAL-2 coding sequence (~1.2 kb) was amplified by nested PCR. ( B ) The AAL-2 coding sequence was cloned into pET-30a plasmids followed by transformation into E. coli BL21 cells. A strong band appeared at approximately 43 kDa after IPTG-induction for 5 h compared with non-induction. ( C ) nAAL-2 and rAAL-2 showed the same electrophoretic mobility on SDS/PAGE and shared the same immunogenicity after immunoblotting with the anti-nAAL-2 polyclonal antibody. Molecular masses are indicated in kDa.
    Plant Rna Isolation Reagent, supplied by Bioteke Corporation, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Promega sv total rna isolation reagent
    Muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts . A. <t>RT-PCR</t> assays were performed with <t>RNA</t> preparations from differentiated C2C12 cells (M), using heart (H), liver (L) or brain (B) RNA preparations as positive controls. The primers used are indicated in the Method section. Note that muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts. B. Relative mRNA expression levels of GATA transcripts were evaluated by quantitative real-time PCR, using cyclophilin A as an internal standard.
    Sv Total Rna Isolation Reagent, supplied by Promega, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Molecular Research Center inc rna dna protein isolation tri reagent
    Switching of actinin splicing by overexpression of CELF proteins. ( A ) Western blot analysis for overexpressed CELF proteins in L cells transiently transfected with expression vectors for ETR3, CUG-BP, CELF4, or mock-transfected (MOCK). Proteins were detected using AntiXpress-HRP conjugated antibody (Invitrogen) against the N-terminal AntiXpress tag in the expression plasmid. ( B–D ) L cells were transiently transfected with 0.2 μg of the pA ( B ), pNM ( C ) or pSM ( D ) α-actinin reporters together with 0.8 μg of pGEM4Z (4Z), ETR3, CUG-BP, CELF4, or β-gal expression constructs. In the right-hand section of D , 0.1 μg of expression plasmid for PTB4 was also cotransfected. <t>RNA</t> was harvested after 48 h, and RT-PCR analysis was performed to determine the extent of NM exon skipping and SM exon inclusion. In MOCK lanes, no <t>DNA</t> was transfected into the cells. Values below each lane give the percentage of EF1a-EF2 or EF1a-SM-EF2 splicing and represent mean ± SD for three experiments, except in the experiment with PTB4 cotransfection ( D, right panel) where they represent the values for the experiment shown. In each case, the percentage of each splice product was calculated as a total of all (two or three) possible spliced products in the lane. The data show that ETR3 and CUG-BP promote SM exon inclusion and NM exon skipping, whereas CELF4 causes skipping of both NM and SM exons.
    Rna Dna Protein Isolation Tri Reagent, supplied by Molecular Research Center inc, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Total RNA was extracted from the aorta of different groups using TRIzol reagent; MiR-1 expression was determined using the miRNA plate assay kit; for normalized RNA content, the U6 snRNA was the internal control. a miR-1 expression was detected in control and AS mice. b miR-1 treatment influences miR-1 expression in the aorta. Levels of miR-1 were detected in different miR-treated AS mice, * P

    Journal: Molecular and Cellular Biochemistry

    Article Title: MicroRNA-1 prevents high-fat diet-induced endothelial permeability in apoE knock-out mice

    doi: 10.1007/s11010-013-1606-x

    Figure Lengend Snippet: Total RNA was extracted from the aorta of different groups using TRIzol reagent; MiR-1 expression was determined using the miRNA plate assay kit; for normalized RNA content, the U6 snRNA was the internal control. a miR-1 expression was detected in control and AS mice. b miR-1 treatment influences miR-1 expression in the aorta. Levels of miR-1 were detected in different miR-treated AS mice, * P

    Article Snippet: Total RNA was extracted from the aorta of different groups using Total RNA Isolation Reagent (TRIzol reagent, Invitrogen).

    Techniques: Expressing, Mouse Assay

    Total RNA and mRNA analysis. RT-PCR was performed on total RNA extracted from CAL27 (a) and SCC25 (b) cells at 24 h after CE or GSE administration; no significant changes were detected at 4 h (data not shown). Relative endpoint (RE) RT-PCR revealed that CE and GSE treatment increased expression of apoptosis-related mRNA (caspase-2, caspase-8) in both cell lines, which is first detectable at 24 h. CE significantly enhanced CAL27 expression of cell-cycle genes (p53, c-myc, ODC), while GSE reduced SCC25 expression of these same targets at this time point.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Cranberry and Grape Seed Extracts Inhibit the Proliferative Phenotype of Oral Squamous Cell Carcinomas

    doi: 10.1093/ecam/nen047

    Figure Lengend Snippet: Total RNA and mRNA analysis. RT-PCR was performed on total RNA extracted from CAL27 (a) and SCC25 (b) cells at 24 h after CE or GSE administration; no significant changes were detected at 4 h (data not shown). Relative endpoint (RE) RT-PCR revealed that CE and GSE treatment increased expression of apoptosis-related mRNA (caspase-2, caspase-8) in both cell lines, which is first detectable at 24 h. CE significantly enhanced CAL27 expression of cell-cycle genes (p53, c-myc, ODC), while GSE reduced SCC25 expression of these same targets at this time point.

    Article Snippet: RNA was isolated from 1.5 × 107 cells of CAL27 and SCC25 cells at 24 h after CE or GSE administration using ABgene Total RNA Isolation Reagent (Epsom, Surrey, UK) and the procedure recommended by the manufacturer.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

    Total RNA and DNA analysis of CAL 27 cells . RT-PCR performed on total RNA extracted from OSCC, CAL 27, revealed that CAL 27 did not express HPV 16-specific mRNA (A: Lane 1). Mock transfectants (A: Lane 2) also did not express HPV. HPV 16-transfected cells (A: Lane 3) expressed HPV 16; specific primers were tested using PCR and the full-length HPV 16-DNA template (A: Lane 4). Total DNA isolated from CAL 27 cells (B: Lane 1) and CAL 27-TF16 cells (B: Lane 3) demonstrated DNA alterations following PAC treatment (50 μg/mL), including a more diffuse banding pattern and lower molecular weight, possibly indicating the onset of DNA fragmentation among the treated cells (B: Lane 2, 4).

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Oral squamous cell carcinoma proliferative phenotype is modulated by proanthocyanidins: a potential prevention and treatment alternative for oral cancer

    doi: 10.1186/1472-6882-7-22

    Figure Lengend Snippet: Total RNA and DNA analysis of CAL 27 cells . RT-PCR performed on total RNA extracted from OSCC, CAL 27, revealed that CAL 27 did not express HPV 16-specific mRNA (A: Lane 1). Mock transfectants (A: Lane 2) also did not express HPV. HPV 16-transfected cells (A: Lane 3) expressed HPV 16; specific primers were tested using PCR and the full-length HPV 16-DNA template (A: Lane 4). Total DNA isolated from CAL 27 cells (B: Lane 1) and CAL 27-TF16 cells (B: Lane 3) demonstrated DNA alterations following PAC treatment (50 μg/mL), including a more diffuse banding pattern and lower molecular weight, possibly indicating the onset of DNA fragmentation among the treated cells (B: Lane 2, 4).

    Article Snippet: RNA was isolated from 1.5 × 107 cells of CAL 27, CAL 27-mTF, and CAL 27-TF16, using ABgene Total RNA Isolation Reagent (Epsom, Surrey, UK) and the procedure recommended by the manufacture.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Polymerase Chain Reaction, Isolation, Molecular Weight

    Custom microarray analysis of the accumulation of antisense RNAs in wild-type and rdr1/2/6 plants subjected to drought stress. A, MA plots illustrating the accumulation level of poly (A-) sense RNA and poly (A-) antisense RNA at 7,138 gene loci in which

    Journal:

    Article Title: Novel Stress-Inducible Antisense RNAs of Protein-Coding Loci Are Synthesized by RNA-Dependent RNA Polymerase

    doi: 10.1104/pp.17.00787

    Figure Lengend Snippet: Custom microarray analysis of the accumulation of antisense RNAs in wild-type and rdr1/2/6 plants subjected to drought stress. A, MA plots illustrating the accumulation level of poly (A-) sense RNA and poly (A-) antisense RNA at 7,138 gene loci in which

    Article Snippet: Total RNAs were extracted with a Plant RNA Isolation Reagent (Life Technologies) and treated with DNase I (Life Technologies).

    Techniques: Microarray

    Schematic model for antisense RNA-mediated degradation of sense mRNAs under abiotic stress. A, The hypothetical model is constructed from the current study and previous reports. Antisense RNAs are generated from uncapped sense RNAs by RDRs. The antisense

    Journal:

    Article Title: Novel Stress-Inducible Antisense RNAs of Protein-Coding Loci Are Synthesized by RNA-Dependent RNA Polymerase

    doi: 10.1104/pp.17.00787

    Figure Lengend Snippet: Schematic model for antisense RNA-mediated degradation of sense mRNAs under abiotic stress. A, The hypothetical model is constructed from the current study and previous reports. Antisense RNAs are generated from uncapped sense RNAs by RDRs. The antisense

    Article Snippet: Total RNAs were extracted with a Plant RNA Isolation Reagent (Life Technologies) and treated with DNase I (Life Technologies).

    Techniques: Construct, Generated

    Analysis of the accumulation of sense and antisense RNA at the RD29A locus in rdr and nrpd1 mutants. A, Schematic diagram of two types of antisense RNAs, fAsRD29A1 and fAsRD29A2 , that exist at the RD29A locus. In order to detect the accumulation of fAsRD29A1

    Journal:

    Article Title: Novel Stress-Inducible Antisense RNAs of Protein-Coding Loci Are Synthesized by RNA-Dependent RNA Polymerase

    doi: 10.1104/pp.17.00787

    Figure Lengend Snippet: Analysis of the accumulation of sense and antisense RNA at the RD29A locus in rdr and nrpd1 mutants. A, Schematic diagram of two types of antisense RNAs, fAsRD29A1 and fAsRD29A2 , that exist at the RD29A locus. In order to detect the accumulation of fAsRD29A1

    Article Snippet: Total RNAs were extracted with a Plant RNA Isolation Reagent (Life Technologies) and treated with DNase I (Life Technologies).

    Techniques:

    Relative expression levels of selected genes as determined by real time RT-PCR. Real time RT-PCR was performed for selected genes of the stress response (A) and genes of RNA processing and degradation (B). Cells were treated by blue light illumination or 1 O 2 exposure and RNA was isolated as described. To visualize gene induction or repression RNA was also isolated of untreated cells. White bars depict the R. sphaeroides wild type after 60 min blue light treatment compared to untreated cells. Grey bars show the results for the cryB deletion mutant under the same conditions. White, striped bars correspond to wild type treated by 20 min 1 O 2 compared to unstressed cells. Grey, striped bars show the results for the cryB deletion under photooxidative stress. Gene annotations are the same as listed in Table 1 or Figure 2 .

    Journal: PLoS ONE

    Article Title: Effects of the Cryptochrome CryB from Rhodobacter sphaeroides on Global Gene Expression in the Dark or Blue Light or in the Presence of Singlet Oxygen

    doi: 10.1371/journal.pone.0033791

    Figure Lengend Snippet: Relative expression levels of selected genes as determined by real time RT-PCR. Real time RT-PCR was performed for selected genes of the stress response (A) and genes of RNA processing and degradation (B). Cells were treated by blue light illumination or 1 O 2 exposure and RNA was isolated as described. To visualize gene induction or repression RNA was also isolated of untreated cells. White bars depict the R. sphaeroides wild type after 60 min blue light treatment compared to untreated cells. Grey bars show the results for the cryB deletion mutant under the same conditions. White, striped bars correspond to wild type treated by 20 min 1 O 2 compared to unstressed cells. Grey, striped bars show the results for the cryB deletion under photooxidative stress. Gene annotations are the same as listed in Table 1 or Figure 2 .

    Article Snippet: For real time RT-PCR experiments total RNA was isolated by Total RNA isolation reagent (TRIR, ABGene) according to the manufacturer's specifications.

    Techniques: Expressing, Quantitative RT-PCR, Isolation, Mutagenesis

    Expression ratio of selected genes as determined by real time RT-PCR. Cells were treated and total RNA was isolated and prepared for real time RT-PCR as described. Categories are clustered as described in Table 1 , including genes involved in photosynthesis (A), citric acid cycle and oxydative phosphorylation (B), stress response (C), transcriptional regulators (D), RNA degradation and processing (E) and others (F, G). White bars indicate the expression ratio, comparing the cryB deletion mutant to the wild type, after 60 minutes semiaerobic blue light treatment. Grey bars depict the expression in 2.4.1Δ cryB after 20 minutes aerobic singlet oxygen treatment compared to the wild type treated in the same manner. Black bars show the expression ratio of the two strains after non-stressed, microaerobic growth. Numbers correspond to R. sphaeroides gene annotations. If gene name is missing, descriptions can be found in Table 1 .

    Journal: PLoS ONE

    Article Title: Effects of the Cryptochrome CryB from Rhodobacter sphaeroides on Global Gene Expression in the Dark or Blue Light or in the Presence of Singlet Oxygen

    doi: 10.1371/journal.pone.0033791

    Figure Lengend Snippet: Expression ratio of selected genes as determined by real time RT-PCR. Cells were treated and total RNA was isolated and prepared for real time RT-PCR as described. Categories are clustered as described in Table 1 , including genes involved in photosynthesis (A), citric acid cycle and oxydative phosphorylation (B), stress response (C), transcriptional regulators (D), RNA degradation and processing (E) and others (F, G). White bars indicate the expression ratio, comparing the cryB deletion mutant to the wild type, after 60 minutes semiaerobic blue light treatment. Grey bars depict the expression in 2.4.1Δ cryB after 20 minutes aerobic singlet oxygen treatment compared to the wild type treated in the same manner. Black bars show the expression ratio of the two strains after non-stressed, microaerobic growth. Numbers correspond to R. sphaeroides gene annotations. If gene name is missing, descriptions can be found in Table 1 .

    Article Snippet: For real time RT-PCR experiments total RNA was isolated by Total RNA isolation reagent (TRIR, ABGene) according to the manufacturer's specifications.

    Techniques: Expressing, Quantitative RT-PCR, Isolation, Mutagenesis

    DNA and mRNA analysis . A) RT-PCR performed on total RNA extracted from cells at d1 (24 hr) following FA administration [400 μg/mL] revealed little change in p53 mRNA transcription in HGF-1 cells, but decreased transcription in CAL27, SCC15, and SCC25 cells. HPV16 and HPV18 had little effect on FA-induced changes in p53 transcription in CAL27 and SCC25 cells, although decreases were observed in both SCC15 and HGF-1 cells. B) PCR screening from DNA confirmed HPV16 and HPV18 DNA was present in the control cell lines, CaSKi (HPV16) and GH354 (HPV18) cervical adenocarcinomas, but not HGF-1, SCC25, SCC15 or CAL27. RT-PCR screening also confirmed the presence of HPV-specific mRNA in the control, but not experimental, cell lines. C) DNA screening post-transfection confirmed the presence of HPV-specific DNA in CAL27, SCC25, SCC15 and HGF-1, but not mock transfectants (mTF); RNA screening confirmed production of HPV-specific mRNA at levels similar to control cell lines.

    Journal: Cancer Cell International

    Article Title: Folic acid supplementation increases survival and modulates high risk HPV-induced phenotypes in oral squamous cell carcinoma cells and correlates with p53 mRNA transcriptional down-regulation

    doi: 10.1186/1475-2867-12-10

    Figure Lengend Snippet: DNA and mRNA analysis . A) RT-PCR performed on total RNA extracted from cells at d1 (24 hr) following FA administration [400 μg/mL] revealed little change in p53 mRNA transcription in HGF-1 cells, but decreased transcription in CAL27, SCC15, and SCC25 cells. HPV16 and HPV18 had little effect on FA-induced changes in p53 transcription in CAL27 and SCC25 cells, although decreases were observed in both SCC15 and HGF-1 cells. B) PCR screening from DNA confirmed HPV16 and HPV18 DNA was present in the control cell lines, CaSKi (HPV16) and GH354 (HPV18) cervical adenocarcinomas, but not HGF-1, SCC25, SCC15 or CAL27. RT-PCR screening also confirmed the presence of HPV-specific mRNA in the control, but not experimental, cell lines. C) DNA screening post-transfection confirmed the presence of HPV-specific DNA in CAL27, SCC25, SCC15 and HGF-1, but not mock transfectants (mTF); RNA screening confirmed production of HPV-specific mRNA at levels similar to control cell lines.

    Article Snippet: To determine the cellular phenotype of p53 mRNA transcription, as well as to confirm HPV-specific mRNA production post-transfection, RNA was isolated from 1.5 × 107 cells after HPV transfection and also following FA administration using ABgene Total RNA Isolation Reagent (Epsom: Surrey, UK) and the procedure recommended by the manufacturer.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Transfection

    Temporal expression of a poCI mRNA. RT-PCR of RNA isolated from whole embryos between stage 4 and stage 32. Chordin serves as a staging control and Ornithine decarboxylase (ODC) as a control for input RNA levels.

    Journal: PLoS ONE

    Article Title: Cloning and spatiotemporal expression of Xenopus laevis Apolipoprotein CI

    doi: 10.1371/journal.pone.0191470

    Figure Lengend Snippet: Temporal expression of a poCI mRNA. RT-PCR of RNA isolated from whole embryos between stage 4 and stage 32. Chordin serves as a staging control and Ornithine decarboxylase (ODC) as a control for input RNA levels.

    Article Snippet: Total RNA was prepared using RNA Bee RNA isolation reagent from Tel-Test Inc. RT-PCR was performed as described [ ].

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    Co-culture of hMSCs transduced with ADAMTS5-targeting shRNA effectively reduces ADAMTS5 expression in flow cytometry sorted RFP-labeled SW982. (A) Following co-culture of non-targeting control shRNA or ADAMTS5-targeting shRNA transduced hMSCs with RFP labeled SW982 cells, the cells were sorted by flow cytometry. The gating threshold (RFP+ sorted) was set based on the analysis of co-cultured hMSCs with unlabeled SW982 cells (no SW982-RFP cells). RFP-positive SW982 cells that were co-cultured with hMSCs previously transduced with non-targeting control-shRNA or ADAMTS5-targeting shRNA were sorted directly into TriPure reagent for RNA isolation. (B) The histogram shows the relative gene expression for ADAMTS5 in the RFP-positive cell population isolated after co-culture with hMSC-shRNA-control or hMSC-shRNA-ADAMTS5 cells as determined by quantitative real time RT-PCR. The result is from a representative experiment.

    Journal: PLoS ONE

    Article Title: Connexin43 Mediated Delivery of ADAMTS5 Targeting siRNAs from Mesenchymal Stem Cells to Synovial Fibroblasts

    doi: 10.1371/journal.pone.0129999

    Figure Lengend Snippet: Co-culture of hMSCs transduced with ADAMTS5-targeting shRNA effectively reduces ADAMTS5 expression in flow cytometry sorted RFP-labeled SW982. (A) Following co-culture of non-targeting control shRNA or ADAMTS5-targeting shRNA transduced hMSCs with RFP labeled SW982 cells, the cells were sorted by flow cytometry. The gating threshold (RFP+ sorted) was set based on the analysis of co-cultured hMSCs with unlabeled SW982 cells (no SW982-RFP cells). RFP-positive SW982 cells that were co-cultured with hMSCs previously transduced with non-targeting control-shRNA or ADAMTS5-targeting shRNA were sorted directly into TriPure reagent for RNA isolation. (B) The histogram shows the relative gene expression for ADAMTS5 in the RFP-positive cell population isolated after co-culture with hMSC-shRNA-control or hMSC-shRNA-ADAMTS5 cells as determined by quantitative real time RT-PCR. The result is from a representative experiment.

    Article Snippet: RFP-positive cells were sorted directly into TriPure RNA isolation reagent (Roche, Indianapolis, IN) for subsequent RNA isolation, reverse transcription and real time PCR.

    Techniques: Co-Culture Assay, Transduction, shRNA, Expressing, Flow Cytometry, Cytometry, Labeling, Cell Culture, Isolation, Quantitative RT-PCR

    Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR. A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1 - RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.

    Journal: PLoS ONE

    Article Title: AaERF1 Positively Regulates the Resistance to Botrytis cinerea in Artemisia annua

    doi: 10.1371/journal.pone.0057657

    Figure Lengend Snippet: Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR. A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1 - RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.

    Article Snippet: Different tissues of A. annua and Arabidopsis plants were collected for RNA extraction using plant RNA isolation reagent (Tiangen Biotech, Beijing) following the manufacturer’s instructions.

    Techniques: Expressing, Polymerase Chain Reaction, Isolation

    MARCH2 expression is required for carvedilol-induced ubiquitination, lysosomal trafficking, and degradation of endogenous β 2 ARs. (A) Rat VSMCs were transfected with either control (CTL) or MARCH2 siRNA; total RNA was extracted 48 h later and subjected to RT-PCR for MARCH2 or GAPDH, as indicated. There was an 80 ± 5% knockdown of MARCH2 mRNA ( n = 6). (B) Rat VSMCs transfected with CTL or MARCH2 siRNA were stimulated with carvedilol (1 h, 37°C), and β 2 AR immunoprecipitates were probed with anti-Ub (top) or anti-β 2 AR (bottom) IgG. (C) The ubiquitin signals were quantified, normalized to the β 2 ARs in each IP sample, and plotted as bar graphs (mean ± SEM). Compared with NS: *, P

    Journal: The Journal of Cell Biology

    Article Title: MARCH2 promotes endocytosis and lysosomal sorting of carvedilol-bound ?2-adrenergic receptors

    doi: 10.1083/jcb.201208192

    Figure Lengend Snippet: MARCH2 expression is required for carvedilol-induced ubiquitination, lysosomal trafficking, and degradation of endogenous β 2 ARs. (A) Rat VSMCs were transfected with either control (CTL) or MARCH2 siRNA; total RNA was extracted 48 h later and subjected to RT-PCR for MARCH2 or GAPDH, as indicated. There was an 80 ± 5% knockdown of MARCH2 mRNA ( n = 6). (B) Rat VSMCs transfected with CTL or MARCH2 siRNA were stimulated with carvedilol (1 h, 37°C), and β 2 AR immunoprecipitates were probed with anti-Ub (top) or anti-β 2 AR (bottom) IgG. (C) The ubiquitin signals were quantified, normalized to the β 2 ARs in each IP sample, and plotted as bar graphs (mean ± SEM). Compared with NS: *, P

    Article Snippet: After transfection with siRNA, cells were harvested in 1 ml Ultraspec RNA, total RNA isolation reagent (Biotex Laboratories, Inc.), and prepared by chloroform extraction and isopropanol precipitation.

    Techniques: Expressing, Transfection, CTL Assay, Reverse Transcription Polymerase Chain Reaction

    Northern blot analysis examining gene expression in cells infected with wild-type (WT) or STOP-mutant EHV-1. RK13 cells were infected with wild-type EHV-1 or the UL4aa18stop EHV-1, and RNA was isolated at 4, 8, and 12 hours post-infection. Northern blot

    Journal:

    Article Title: The UL4 protein of equine herpesvirus 1 is not essential for replication or pathogenesis and inhibits gene expression controlled by viral and heterologous promoters

    doi: 10.1016/j.virol.2011.01.025

    Figure Lengend Snippet: Northern blot analysis examining gene expression in cells infected with wild-type (WT) or STOP-mutant EHV-1. RK13 cells were infected with wild-type EHV-1 or the UL4aa18stop EHV-1, and RNA was isolated at 4, 8, and 12 hours post-infection. Northern blot

    Article Snippet: Northern blot analysis was performed by isolating total RNA from RK13 cells infected with RacL11 EHV-1 at the indicated times using the RNA-Bee RNA isolation reagent (AMS Biotechnology (Europe) Ltd, Abingdon, U.K.) per the manufacturer’s procedure ( ).

    Techniques: Northern Blot, Expressing, Infection, Mutagenesis, Isolation

    Small interfering RNA specific for PIP5KI β (but not α or γ ) reduces PIP2 levels in HeLa cells. HeLa cells were transfected with siRNA oligos targeting the α, β, or γ isoforms of PIP5KI or an irrelevant sequence (control, C). Western blots with antibodies specific for the respective protein and tubulin (loading control) were performed 48 h after transfection. (A) An example of a Western blot specific for PIP5KIα is shown. Similar blots were quantified by densitometry, and the values are presented graphically. (B) PIP2 levels in cultures transfected with siRNAs specific for the enzymes shown were measured as described in Fig. 1 B. (C) mRNA for each enzyme in cells treated with siRNA specific for one isoform were measured by real-time PCR. For each isoform, values in A and B are expressed as a percentage of the value measured in the control transfected with unrelated siRNA. All graphs present the averages of at least three experiments with SD.

    Journal: The Journal of Cell Biology

    Article Title: Phosphatidylinositol phosphate 5-kinase I? recruits AP-2 to the plasma membrane and regulates rates of constitutive endocytosis

    doi: 10.1083/jcb.200302051

    Figure Lengend Snippet: Small interfering RNA specific for PIP5KI β (but not α or γ ) reduces PIP2 levels in HeLa cells. HeLa cells were transfected with siRNA oligos targeting the α, β, or γ isoforms of PIP5KI or an irrelevant sequence (control, C). Western blots with antibodies specific for the respective protein and tubulin (loading control) were performed 48 h after transfection. (A) An example of a Western blot specific for PIP5KIα is shown. Similar blots were quantified by densitometry, and the values are presented graphically. (B) PIP2 levels in cultures transfected with siRNAs specific for the enzymes shown were measured as described in Fig. 1 B. (C) mRNA for each enzyme in cells treated with siRNA specific for one isoform were measured by real-time PCR. For each isoform, values in A and B are expressed as a percentage of the value measured in the control transfected with unrelated siRNA. All graphs present the averages of at least three experiments with SD.

    Article Snippet: Total RNA was extracted from HeLa cells transfected with siRNAs using the total RNA/mRNA isolation reagent RNA STAT-60™ (TEL-TEST, Inc.).

    Techniques: Small Interfering RNA, Transfection, Sequencing, Western Blot, Real-time Polymerase Chain Reaction

    Expression of mouse Dlx4 mRNA in E10.5 yolk sac-derived blood, E13.5 fetal liver, adult bone marrow, and adult reticulocytes. One µg of total RNA of mouse yolk sac blood, fetal liver blood, adult bone marrow (BM), and adult reticulocytes was subjected

    Journal:

    Article Title: BP1 motif in the human ?-globin promoter affects ?-globin expression during embryonic/fetal erythropoiesis in transgenic mice bearing the human ?-globin gene

    doi: 10.1016/j.bcmd.2008.05.008

    Figure Lengend Snippet: Expression of mouse Dlx4 mRNA in E10.5 yolk sac-derived blood, E13.5 fetal liver, adult bone marrow, and adult reticulocytes. One µg of total RNA of mouse yolk sac blood, fetal liver blood, adult bone marrow (BM), and adult reticulocytes was subjected

    Article Snippet: Total RNA was isolated from human or mouse cells using RNA STAT-60 total RNA/mRNA isolation reagent (Tel-Test, Inc., Friendswood, TX) according to the manufacturer’s protocol and quantitated by absorbance at 260 nm.

    Techniques: Expressing, Derivative Assay

    Cloning, expression and purification of AAL-2 ( A ) Total RNA was isolated from fresh A. aegerita fruiting body and amplified by reverse transcription–PCR. The AAL-2 coding sequence (~1.2 kb) was amplified by nested PCR. ( B ) The AAL-2 coding sequence was cloned into pET-30a plasmids followed by transformation into E. coli BL21 cells. A strong band appeared at approximately 43 kDa after IPTG-induction for 5 h compared with non-induction. ( C ) nAAL-2 and rAAL-2 showed the same electrophoretic mobility on SDS/PAGE and shared the same immunogenicity after immunoblotting with the anti-nAAL-2 polyclonal antibody. Molecular masses are indicated in kDa.

    Journal: Biochemical Journal

    Article Title: A novel lectin from Agrocybe aegerita shows high binding selectivity for terminal N-acetylglucosamine

    doi: 10.1042/BJ20112061

    Figure Lengend Snippet: Cloning, expression and purification of AAL-2 ( A ) Total RNA was isolated from fresh A. aegerita fruiting body and amplified by reverse transcription–PCR. The AAL-2 coding sequence (~1.2 kb) was amplified by nested PCR. ( B ) The AAL-2 coding sequence was cloned into pET-30a plasmids followed by transformation into E. coli BL21 cells. A strong band appeared at approximately 43 kDa after IPTG-induction for 5 h compared with non-induction. ( C ) nAAL-2 and rAAL-2 showed the same electrophoretic mobility on SDS/PAGE and shared the same immunogenicity after immunoblotting with the anti-nAAL-2 polyclonal antibody. Molecular masses are indicated in kDa.

    Article Snippet: Total RNA was prepared from the fresh fruiting bodies of A. aegerita by grinding in liquid nitrogen followed by extraction with Plant RNA Isolation Reagent (BioTeke).

    Techniques: Clone Assay, Expressing, Purification, Isolation, Amplification, Polymerase Chain Reaction, Sequencing, Nested PCR, Positron Emission Tomography, Transformation Assay, SDS Page

    Muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts . A. RT-PCR assays were performed with RNA preparations from differentiated C2C12 cells (M), using heart (H), liver (L) or brain (B) RNA preparations as positive controls. The primers used are indicated in the Method section. Note that muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts. B. Relative mRNA expression levels of GATA transcripts were evaluated by quantitative real-time PCR, using cyclophilin A as an internal standard.

    Journal: BMC Molecular Biology

    Article Title: GATA elements control repression of cardiac troponin I promoter activity in skeletal muscle cells

    doi: 10.1186/1471-2199-8-78

    Figure Lengend Snippet: Muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts . A. RT-PCR assays were performed with RNA preparations from differentiated C2C12 cells (M), using heart (H), liver (L) or brain (B) RNA preparations as positive controls. The primers used are indicated in the Method section. Note that muscle cells express GATA-2, -3 and -4 but not GATA-1, -5 and -6 transcripts. B. Relative mRNA expression levels of GATA transcripts were evaluated by quantitative real-time PCR, using cyclophilin A as an internal standard.

    Article Snippet: For real-time PCR assays, total RNA was prepared from C2C12 myotubes with SV Total RNA Isolation reagent (Promega).

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

    Switching of actinin splicing by overexpression of CELF proteins. ( A ) Western blot analysis for overexpressed CELF proteins in L cells transiently transfected with expression vectors for ETR3, CUG-BP, CELF4, or mock-transfected (MOCK). Proteins were detected using AntiXpress-HRP conjugated antibody (Invitrogen) against the N-terminal AntiXpress tag in the expression plasmid. ( B–D ) L cells were transiently transfected with 0.2 μg of the pA ( B ), pNM ( C ) or pSM ( D ) α-actinin reporters together with 0.8 μg of pGEM4Z (4Z), ETR3, CUG-BP, CELF4, or β-gal expression constructs. In the right-hand section of D , 0.1 μg of expression plasmid for PTB4 was also cotransfected. RNA was harvested after 48 h, and RT-PCR analysis was performed to determine the extent of NM exon skipping and SM exon inclusion. In MOCK lanes, no DNA was transfected into the cells. Values below each lane give the percentage of EF1a-EF2 or EF1a-SM-EF2 splicing and represent mean ± SD for three experiments, except in the experiment with PTB4 cotransfection ( D, right panel) where they represent the values for the experiment shown. In each case, the percentage of each splice product was calculated as a total of all (two or three) possible spliced products in the lane. The data show that ETR3 and CUG-BP promote SM exon inclusion and NM exon skipping, whereas CELF4 causes skipping of both NM and SM exons.

    Journal:

    Article Title: Antagonistic regulation of ?-actinin alternative splicing by CELF proteins and polypyrimidine tract binding protein

    doi: 10.1261/rna.2191903

    Figure Lengend Snippet: Switching of actinin splicing by overexpression of CELF proteins. ( A ) Western blot analysis for overexpressed CELF proteins in L cells transiently transfected with expression vectors for ETR3, CUG-BP, CELF4, or mock-transfected (MOCK). Proteins were detected using AntiXpress-HRP conjugated antibody (Invitrogen) against the N-terminal AntiXpress tag in the expression plasmid. ( B–D ) L cells were transiently transfected with 0.2 μg of the pA ( B ), pNM ( C ) or pSM ( D ) α-actinin reporters together with 0.8 μg of pGEM4Z (4Z), ETR3, CUG-BP, CELF4, or β-gal expression constructs. In the right-hand section of D , 0.1 μg of expression plasmid for PTB4 was also cotransfected. RNA was harvested after 48 h, and RT-PCR analysis was performed to determine the extent of NM exon skipping and SM exon inclusion. In MOCK lanes, no DNA was transfected into the cells. Values below each lane give the percentage of EF1a-EF2 or EF1a-SM-EF2 splicing and represent mean ± SD for three experiments, except in the experiment with PTB4 cotransfection ( D, right panel) where they represent the values for the experiment shown. In each case, the percentage of each splice product was calculated as a total of all (two or three) possible spliced products in the lane. The data show that ETR3 and CUG-BP promote SM exon inclusion and NM exon skipping, whereas CELF4 causes skipping of both NM and SM exons.

    Article Snippet: Transiently expressed cytoplasmic RNA was harvested using RNA/DNA/protein isolation TRI-reagent (Molecular Research Centre Inc.), and analyzed by RT-PCR as previously described ( ; ), and quantitated using a Molecular Dynamics Phosphorimager ( ; ).

    Techniques: Over Expression, Western Blot, Transfection, Expressing, Plasmid Preparation, Construct, Reverse Transcription Polymerase Chain Reaction, Cotransfection