rneasy kit Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Qiagen rneasy mini kit
    Slot blot detection of dsRNA in extracts from virus-infected cells. Cells were infected with virus and then total <t>RNA</t> was extracted at multiple time points post-infection using the <t>RNeasy</t> Kit (Qiagen). Equal volumes of extract were then transferred onto
    Rneasy Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 344222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy mini kit/product/Qiagen
    Average 99 stars, based on 344222 article reviews
    Price from $9.99 to $1999.99
    rneasy mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy kit
    Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total <t>RNA</t> was harvested from the peripheral blood mononuclear cells using an <t>RNeasy</t> kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.
    Rneasy Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 115893 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy kit/product/Qiagen
    Average 99 stars, based on 115893 article reviews
    Price from $9.99 to $1999.99
    rneasy kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy plant mini kit
    The efficiency of filter paper for purification of nucleic acids from various sources using respective <t>Qiagen</t> kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen <t>RNeasy</t> plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.
    Rneasy Plant Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 39071 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy plant mini kit/product/Qiagen
    Average 99 stars, based on 39071 article reviews
    Price from $9.99 to $1999.99
    rneasy plant mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy micro kit
    Characterization of endogenous pluripotent makers in selected iPS cell lines. Panel A. Total RNA was isolated using <t>RNeasy</t> Micro Kit from selected iPS cell lines (G1–G3, G5, G6), hES H9 cells (H9), and human primary fibroblasts (F). Total RNA (500 ng) was reverse-transcribed using Superscript III Reverse Transcriptase primed with oligo(dT) 12–18 and used as template in subsequent <t>PCR</t> with Taq DNA Polymerase. PCR analysis examined the expression of endogenous Oct4, Nanog, Sox2, as well as ABCG2, Rex1, DNMT3B and hTERT. GAPDH was used as an internal control. N, no template control (N). PCR products were analyzed on a 10% polyacrylamide TBE Precast Gel. Panel B. TRAP assay for telomerase activity. Selected iPS cells (G1–G3, G6), hES H9 cells (H9), and human primary fibroblasts (F) were analyzed for telomerase activity using the TRAPEZE RT Telomerase Detection Kit as described in M M. PCR products were separated on 10% polyacrylamide TBE Precast Gel. Individual samples are as indicated.
    Rneasy Micro Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 35691 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy micro kit/product/Qiagen
    Average 99 stars, based on 35691 article reviews
    Price from $9.99 to $1999.99
    rneasy micro kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy lipid tissue mini kit
    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at <t>−80°C</t> prior to total RNA preparation using the QIAGEN <t>RNeasy</t> Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.
    Rneasy Lipid Tissue Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 10193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy lipid tissue mini kit/product/Qiagen
    Average 99 stars, based on 10193 article reviews
    Price from $9.99 to $1999.99
    rneasy lipid tissue mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy minelute cleanup kit
    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at <t>−80°C</t> prior to total RNA preparation using the QIAGEN <t>RNeasy</t> Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.
    Rneasy Minelute Cleanup Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 10584 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy minelute cleanup kit/product/Qiagen
    Average 99 stars, based on 10584 article reviews
    Price from $9.99 to $1999.99
    rneasy minelute cleanup kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy midi kit
    Image showing nucleotide size distribution from CTAB (panel A) and CTAB + <t>RNeasy</t> <t>RNA</t> isolations (panel B) . Nucleotides have been resolved by denaturing gel electrophoresis and have been stained using ethidium bromide and viewed under a UV transilluminator. In both samples, clear rRNA bands can be seen, suggesting the integrity of the RNAs. In the CTAB gel, a band of high-molecular weight DNA can be seen, this is absent from the CTAB + RNeasy extractions.
    Rneasy Midi Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 7875 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy midi kit/product/Qiagen
    Average 99 stars, based on 7875 article reviews
    Price from $9.99 to $1999.99
    rneasy midi kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy plus micro kit
    Sorting small cell populations directly into the lysis buffer of the <t>RNA</t> isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the <t>RNeasy</t> plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM
    Rneasy Plus Micro Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 7494 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy plus micro kit/product/Qiagen
    Average 99 stars, based on 7494 article reviews
    Price from $9.99 to $1999.99
    rneasy plus micro kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy fibrous tissue mini kit
    Sorting small cell populations directly into the lysis buffer of the <t>RNA</t> isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the <t>RNeasy</t> plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM
    Rneasy Fibrous Tissue Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 5467 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy fibrous tissue mini kit/product/Qiagen
    Average 99 stars, based on 5467 article reviews
    Price from $9.99 to $1999.99
    rneasy fibrous tissue mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy ffpe kit
    Bland-Altman plots for investigating the level of agreement between RNA extraction kits. Each plot shows the differences between the two kits against the averages of the two kits. The lines represent the mean differences and upper and lower limits of agreement (LOA, mean differences ±1.96SD). a Comparison of RNA yield (ng/μl) of samples extracted with High Pure <t>FFPE</t> RNA Micro Kit and <t>RNeasy®</t> FFPE kit. b Comparison of purity (A260/A280) of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. c Comparison of RIN-values of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. d Comparison of RNA yield (ng/μl) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. e Comparison of purity (A260/A280) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. f Comparison of RIN-values of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit
    Rneasy Ffpe Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 5166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy ffpe kit/product/Qiagen
    Average 99 stars, based on 5166 article reviews
    Price from $9.99 to $1999.99
    rneasy ffpe kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy extraction kit
    The decreased levels of <t>CCR5</t> expression by CCR5 shRNA (1005) correlates with that of CCR5 mRNA . huCCR5-293T cells were transduced with lentiviral vectors bearing either shRNA 1005 against CCR5 {CCR5shRNA (1005)} or a control shRNA against firefly luciferase (Luc shRNA). To monitor the expression levels of CCR5 on cell surface, the cells were cultured for 4 days and stained with either PE-Cy5 conjugated anti human CCR5 monoclonal antibody or isotype control. CCR5 and EGFP expression were analyzed by flow cytometry. The percentage number in each quadrant is indicated in each panel (A). To measure the levels of CCR5 mRNA, total RNA was isolated using Qiagen <t>RNeasy</t> extraction kit. Quantitative RT-PCR was performed using IQ5 with iScript one step RT-PCR kit using β-actin as an internal control (B).
    Rneasy Extraction Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 3522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy extraction kit/product/Qiagen
    Average 99 stars, based on 3522 article reviews
    Price from $9.99 to $1999.99
    rneasy extraction kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Qiagen rneasy protect mini kit
    The decreased levels of <t>CCR5</t> expression by CCR5 shRNA (1005) correlates with that of CCR5 mRNA . huCCR5-293T cells were transduced with lentiviral vectors bearing either shRNA 1005 against CCR5 {CCR5shRNA (1005)} or a control shRNA against firefly luciferase (Luc shRNA). To monitor the expression levels of CCR5 on cell surface, the cells were cultured for 4 days and stained with either PE-Cy5 conjugated anti human CCR5 monoclonal antibody or isotype control. CCR5 and EGFP expression were analyzed by flow cytometry. The percentage number in each quadrant is indicated in each panel (A). To measure the levels of CCR5 mRNA, total RNA was isolated using Qiagen <t>RNeasy</t> extraction kit. Quantitative RT-PCR was performed using IQ5 with iScript one step RT-PCR kit using β-actin as an internal control (B).
    Rneasy Protect Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2931 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy protect mini kit/product/Qiagen
    Average 99 stars, based on 2931 article reviews
    Price from $9.99 to $1999.99
    rneasy protect mini kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Slot blot detection of dsRNA in extracts from virus-infected cells. Cells were infected with virus and then total RNA was extracted at multiple time points post-infection using the RNeasy Kit (Qiagen). Equal volumes of extract were then transferred onto

    Journal: Virology

    Article Title: Monkeypox virus induces the synthesis of less dsRNA than vaccinia virus, and is more resistant to the anti-poxvirus drug, IBT, than vaccinia virus

    doi: 10.1016/j.virol.2016.07.016

    Figure Lengend Snippet: Slot blot detection of dsRNA in extracts from virus-infected cells. Cells were infected with virus and then total RNA was extracted at multiple time points post-infection using the RNeasy Kit (Qiagen). Equal volumes of extract were then transferred onto

    Article Snippet: Total RNA was extracted with the RNeasy Mini Kit, using QiaShredder homogenization and in-column DNase treatment as described by the manufacturer (Qiagen).

    Techniques: Dot Blot, Infection

    CD63 mRNA down regulation by siRNA in human MDMs which are challenged with different concentration of HIV-1. A. MDMs (5 × 10 5 cells/well) were plated in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). For controls, cells were treated with AZT (1 mM) or raltegravir (20 mM). Controls also included untreated cells or cells infected with HIV-1 SX (m.o.i. = 0.02) only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. MDMs were challenged with different concentration of HIV-1 SX (m.o.i. = 0.6, 0.2. 0.06 and 0.02, respectively). Supernatants were harvested for p24 detection on day 7 post-infection for MDMs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA).

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Tetraspanin CD63 is a regulator of HIV-1 replication

    doi:

    Figure Lengend Snippet: CD63 mRNA down regulation by siRNA in human MDMs which are challenged with different concentration of HIV-1. A. MDMs (5 × 10 5 cells/well) were plated in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). For controls, cells were treated with AZT (1 mM) or raltegravir (20 mM). Controls also included untreated cells or cells infected with HIV-1 SX (m.o.i. = 0.02) only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. MDMs were challenged with different concentration of HIV-1 SX (m.o.i. = 0.6, 0.2. 0.06 and 0.02, respectively). Supernatants were harvested for p24 detection on day 7 post-infection for MDMs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA).

    Article Snippet: Total mRNA was isolated from siRNA-transfected cells and MDMs using RNeasy Mini Kits (Qiagen).

    Techniques: Concentration Assay, Transfection, Infection, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

    Effects of CD63 silencing on HIV-1 replication in human PBLs and DCs. A. PBLs (1 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. PBLs cells (1 × 10 5 cells/well) or (C) DCs cells (5 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Forty eight hours post-transfection, cells were infected with HIV-1 89.6 (m.o.i. = 0.02). Supernatants were harvested for p24 detection on day 5 post-infection for PBLs cells, and on day 7 post-infection for DCs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA). C. Cell lysates from MDMs on day 7 post-infection were also harvested for detection of intracellular p24. *P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Tetraspanin CD63 is a regulator of HIV-1 replication

    doi:

    Figure Lengend Snippet: Effects of CD63 silencing on HIV-1 replication in human PBLs and DCs. A. PBLs (1 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Total mRNA was isolated from each well using the Qiagen RNeasy kit 48 h after transfection. Quantitative RT-PCR was used to determine relative CD63 expression levels, normalizing to GAPDH expression as an internal control. B. PBLs cells (1 × 10 5 cells/well) or (C) DCs cells (5 × 10 5 cells/well) were plated in triplicate in 24-well plates and transfected with 50 nM siRNAs (CD63, CD4, and ERBB2IP). Controls included untreated cells or cells treated with virus infection only. Forty eight hours post-transfection, cells were infected with HIV-1 89.6 (m.o.i. = 0.02). Supernatants were harvested for p24 detection on day 5 post-infection for PBLs cells, and on day 7 post-infection for DCs using p24 Capture ELISA kit (ImmunoDiagnostics, Woburn, MA). C. Cell lysates from MDMs on day 7 post-infection were also harvested for detection of intracellular p24. *P

    Article Snippet: Total mRNA was isolated from siRNA-transfected cells and MDMs using RNeasy Mini Kits (Qiagen).

    Techniques: Transfection, Infection, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

    Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Therapeutic effects of calcium dobesilate on diabetic nephropathy mediated through reduction of expression of PAI-1

    doi: 10.3892/etm.2012.755

    Figure Lengend Snippet: Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Article Snippet: The total RNA was harvested from peripheral blood mononuclear cells using the RNeasy kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

    Techniques: Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction, Sequencing, Amplification, Polymerase Chain Reaction

    Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Journal: PLoS ONE

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    doi: 10.1371/journal.pone.0214609

    Figure Lengend Snippet: Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Article Snippet: Optimizing RNA isolation using the RNeasy Plus Mini kit The RNeasy Plus Mini kit offers two alternative protocols; the standard protocol provides enrichment of the sample with intact mRNAs by eliminating RNAs shorter than 200 nucleotides; the second protocol is designed for the purification of total RNA.

    Techniques: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Isolation, Molecular Weight, Marker

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Journal: PLoS ONE

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    doi: 10.1371/journal.pone.0214609

    Figure Lengend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Article Snippet: Optimizing RNA isolation using the RNeasy Plus Mini kit The RNeasy Plus Mini kit offers two alternative protocols; the standard protocol provides enrichment of the sample with intact mRNAs by eliminating RNAs shorter than 200 nucleotides; the second protocol is designed for the purification of total RNA.

    Techniques: Real-time Polymerase Chain Reaction, Isolation

    SPP1 expression is up-regulated in glioma initiating cells ( A ) Representative examples of dot plots and histograms of Rhod 123(–) subpopulations from 5 glioma cell lines were acquired by flow cytometry. Glioma cells (1 × 10 7 ) were trypsinized and incubated with 0.1 μg/ml Rhod123 for 20 min in 37°C. After washing the cells were placed in 37°C for 90 min for Rhod123 exclusion in a dark compartment. Cells kept on ice to inhibit exclusion of Rhod123 were used as a positive control for gating in flow cytometry. Fractions of Rhod123(+) and Rhod123(–) cells were sorted using FACS Aria and a left panel shows a gating strategy. ( B ) Analysis of SPP1, OCT3/4 and NANOG gene expression in Rhod 123(–) subpopulations sorted from four human glioma cell lines. Total RNA was isolated from sorted cells using Qiagen RNeasy kit and the levels of SPP1, NANOG and OCT3/4 mRNA were determined by qPCR in Rhod 123(+) and Rhod 123(–) subpopulations; their expression in Rhod 123(+) subpopulations was taken as 1 (a red line). P values were calculated with a t-test and considered significant when *P ≤ 0.05 and * *P ≤ 0.01. ( C ) Quantification of the expression of Spp1, Oct3/4 and Nanog in Rhod 123(–) subpopulations isolated from rat C6 glioma cells versus their levels in Rhod 123(+) subpopulations taken as 1 (a red line); n = 3.

    Journal: Oncotarget

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma

    doi: 10.18632/oncotarget.14092

    Figure Lengend Snippet: SPP1 expression is up-regulated in glioma initiating cells ( A ) Representative examples of dot plots and histograms of Rhod 123(–) subpopulations from 5 glioma cell lines were acquired by flow cytometry. Glioma cells (1 × 10 7 ) were trypsinized and incubated with 0.1 μg/ml Rhod123 for 20 min in 37°C. After washing the cells were placed in 37°C for 90 min for Rhod123 exclusion in a dark compartment. Cells kept on ice to inhibit exclusion of Rhod123 were used as a positive control for gating in flow cytometry. Fractions of Rhod123(+) and Rhod123(–) cells were sorted using FACS Aria and a left panel shows a gating strategy. ( B ) Analysis of SPP1, OCT3/4 and NANOG gene expression in Rhod 123(–) subpopulations sorted from four human glioma cell lines. Total RNA was isolated from sorted cells using Qiagen RNeasy kit and the levels of SPP1, NANOG and OCT3/4 mRNA were determined by qPCR in Rhod 123(+) and Rhod 123(–) subpopulations; their expression in Rhod 123(+) subpopulations was taken as 1 (a red line). P values were calculated with a t-test and considered significant when *P ≤ 0.05 and * *P ≤ 0.01. ( C ) Quantification of the expression of Spp1, Oct3/4 and Nanog in Rhod 123(–) subpopulations isolated from rat C6 glioma cells versus their levels in Rhod 123(+) subpopulations taken as 1 (a red line); n = 3.

    Article Snippet: Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template.

    Techniques: Expressing, Flow Cytometry, Cytometry, Incubation, Positive Control, FACS, Isolation, Real-time Polymerase Chain Reaction

    Replication Factors Are Misregulated in set2 Δ Cells (A) Transcript levels of cdc18 + , cdc22 + , and cdt1 + were established in exponentially growing wild-type and set2 Δ cells. RNA was extracted with Qiagen RNeasy kit and relative transcript levels of cdc18 + , cdc22 + , and cdt1 + were established by RT-qPCR. Error bars represent SD of three biological repeats. The asterisk ( ∗ ) represents significant difference compared with wild-type and set2 Δ. (B) Cdc18, Cdt1, or Cdc22 protein levels of exponentially growing wild-type and set2 Δ cells. Cell extracts were prepared from vegetative cells and processed for western blot. Immunoblots of total cell lysates were probed with PAP or GFP antibody. α-Tubulin is shown as a loading control. (C) cdc18-TAP or cdc18-TAP set2 Δ cells were arrested in G1 by nitrogen starvation and released, and samples were taken at time points indicated and subjected to FACS analysis. (D) In parallel, cdc18-TAP and cdc18-TAP set2 Δ cells were processed for western blotting at indicated times. (E) cdt1-TAP or cdt1-TAP set2 Δ cells were arrested in G1 by nitrogen starvation, released, and samples taken at time points indicated and subjected to FACS analysis. (F) In parallel, cdt1-TAP and cdt1-TAP set2 Δ cells were processed for western blotting at indicated times. (G) A similar experiment to that described in (C) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. (H) A similar experiment to that described in (D) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. Immunoblots of total cell extracts were probed with GFP antibody. α-Tubulin is shown as a loading control.

    Journal: Cell Reports

    Article Title: Set2 Methyltransferase Facilitates DNA Replication and Promotes Genotoxic Stress Responses through MBF-Dependent Transcription

    doi: 10.1016/j.celrep.2017.08.058

    Figure Lengend Snippet: Replication Factors Are Misregulated in set2 Δ Cells (A) Transcript levels of cdc18 + , cdc22 + , and cdt1 + were established in exponentially growing wild-type and set2 Δ cells. RNA was extracted with Qiagen RNeasy kit and relative transcript levels of cdc18 + , cdc22 + , and cdt1 + were established by RT-qPCR. Error bars represent SD of three biological repeats. The asterisk ( ∗ ) represents significant difference compared with wild-type and set2 Δ. (B) Cdc18, Cdt1, or Cdc22 protein levels of exponentially growing wild-type and set2 Δ cells. Cell extracts were prepared from vegetative cells and processed for western blot. Immunoblots of total cell lysates were probed with PAP or GFP antibody. α-Tubulin is shown as a loading control. (C) cdc18-TAP or cdc18-TAP set2 Δ cells were arrested in G1 by nitrogen starvation and released, and samples were taken at time points indicated and subjected to FACS analysis. (D) In parallel, cdc18-TAP and cdc18-TAP set2 Δ cells were processed for western blotting at indicated times. (E) cdt1-TAP or cdt1-TAP set2 Δ cells were arrested in G1 by nitrogen starvation, released, and samples taken at time points indicated and subjected to FACS analysis. (F) In parallel, cdt1-TAP and cdt1-TAP set2 Δ cells were processed for western blotting at indicated times. (G) A similar experiment to that described in (C) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. (H) A similar experiment to that described in (D) was carried out using cdc22-CFP or cdc22-CFP set2 Δ cells. Immunoblots of total cell extracts were probed with GFP antibody. α-Tubulin is shown as a loading control.

    Article Snippet: Total RNA was prepared using the RNeasy Plus Kit (Qiagen) as indicated in the manufacturer’s manual.

    Techniques: Quantitative RT-PCR, Western Blot, FACS

    MUC5B-AS1 increases the stability of MUC5B mRNA by forming a protective RNA duplex. a Schematic representation of the PCR amplification regions for overlapping (OL) and non-overlapping (non-OL) regions of MUC5B. We designed two pairs of primers to amplify the OL regions (OL1 and OL2) and non-OL (non-OL1 and non-OL2) regions of MUC5B, respectively. F forward primer, R reverse primer. b RT-PCR products of OL and non-OL regions of MUC5B. Total RNA samples were treated with RNAse A + T cocktail and then cleaned up RNA using RNeasy kits. RT-PCR was conducted using the primers to detect the OL and non-OL regions of the MUC5B mRNA. OL and non-OL regions of KRT7-AS were used as a positive control. c Stability of MUC5B mRNA over 12 h was measured by qRT-PCR relative to time 0 h after blocking new RNA synthesis with Actinomycin D (1 μg/mL; indicated with black arrow). H1299 cells with MUC5B-AS1 or empty vector stable expression were treated with 1 μg/mL ActD, and then harvested cells for RNA purification at 12 h after addition of ActD. Then, MUC5B mRNA stability were subsequently measured by qRT-PCR and were normalized against a synthesized exogenous reference λ polyA + RNA. Student’s t -test, * P

    Journal: Cell Death & Disease

    Article Title: Long non-coding RNA MUC5B-AS1 promotes metastasis through mutually regulating MUC5B expression in lung adenocarcinoma

    doi: 10.1038/s41419-018-0472-6

    Figure Lengend Snippet: MUC5B-AS1 increases the stability of MUC5B mRNA by forming a protective RNA duplex. a Schematic representation of the PCR amplification regions for overlapping (OL) and non-overlapping (non-OL) regions of MUC5B. We designed two pairs of primers to amplify the OL regions (OL1 and OL2) and non-OL (non-OL1 and non-OL2) regions of MUC5B, respectively. F forward primer, R reverse primer. b RT-PCR products of OL and non-OL regions of MUC5B. Total RNA samples were treated with RNAse A + T cocktail and then cleaned up RNA using RNeasy kits. RT-PCR was conducted using the primers to detect the OL and non-OL regions of the MUC5B mRNA. OL and non-OL regions of KRT7-AS were used as a positive control. c Stability of MUC5B mRNA over 12 h was measured by qRT-PCR relative to time 0 h after blocking new RNA synthesis with Actinomycin D (1 μg/mL; indicated with black arrow). H1299 cells with MUC5B-AS1 or empty vector stable expression were treated with 1 μg/mL ActD, and then harvested cells for RNA purification at 12 h after addition of ActD. Then, MUC5B mRNA stability were subsequently measured by qRT-PCR and were normalized against a synthesized exogenous reference λ polyA + RNA. Student’s t -test, * P

    Article Snippet: We then cleaned up RNA using RNeasy kits (QIAGEN, Madison, WI, USA) and carried out reverse transcription-polymerase chain reaction (RT-PCR) using the primers to detect the OL and non-OL regions of the MUC5B mRNA.

    Techniques: Polymerase Chain Reaction, Amplification, Reverse Transcription Polymerase Chain Reaction, Positive Control, Quantitative RT-PCR, Blocking Assay, Plasmid Preparation, Expressing, Purification, Synthesized

    Induction of cytokine genes by POVPC and its metabolites. Time-dependent changes in the expression of (A) TNF-α, (B) IL-8, (C) GM-CSF, (D) IL-1β, (E) MCP-1, and (F) IL-6 in THP-DM treated with 10 μM each of POVPC, lyso-PC, and PHVPC. (G) Changes in cytokine gene expression in THP-DM treated with 10 μM of POVPE or PHVPE for 3 h. The THP-1 cells were differentiated with 100 nM PMA for 72 h, incubated in media with 0.1% FBS for an additional 24 h, and then either left untreated or treated in HBSS with POVPC or POVPE and their metabolites. At indicated times, the cells were harvested, and their total RNA was extracted with RNeasy® Mini Kit (Qiagen). Changes in the levels of mRNA of cytokine genes were measured by quantitative RT-PCR as described in Materials and Methods . Data are mean ± SEM; n = 3. * P

    Journal: Journal of Lipid Research

    Article Title: Reductive metabolism increases the proinflammatory activity of aldehyde phospholipids [S]

    doi: 10.1194/jlr.M013854

    Figure Lengend Snippet: Induction of cytokine genes by POVPC and its metabolites. Time-dependent changes in the expression of (A) TNF-α, (B) IL-8, (C) GM-CSF, (D) IL-1β, (E) MCP-1, and (F) IL-6 in THP-DM treated with 10 μM each of POVPC, lyso-PC, and PHVPC. (G) Changes in cytokine gene expression in THP-DM treated with 10 μM of POVPE or PHVPE for 3 h. The THP-1 cells were differentiated with 100 nM PMA for 72 h, incubated in media with 0.1% FBS for an additional 24 h, and then either left untreated or treated in HBSS with POVPC or POVPE and their metabolites. At indicated times, the cells were harvested, and their total RNA was extracted with RNeasy® Mini Kit (Qiagen). Changes in the levels of mRNA of cytokine genes were measured by quantitative RT-PCR as described in Materials and Methods . Data are mean ± SEM; n = 3. * P

    Article Snippet: RNeasy RNA isolation kit was from Qiagen (Valencia, CA), rDNase (DNA-freeTM Kit) was from Ambion (Austin, TX), AMV Reverse Transcriptase from Promega (Madison, WI), and SYBR Green master mix from Quanta BioSciences (Gaithersburg, MD).

    Techniques: Expressing, Incubation, Quantitative RT-PCR

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Journal: PLoS ONE

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    doi: 10.1371/journal.pone.0203011

    Figure Lengend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook.

    Techniques: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Journal: PLoS ONE

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    doi: 10.1371/journal.pone.0203011

    Figure Lengend Snippet: Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook.

    Techniques: Purification, Agarose Gel Electrophoresis, Amplification, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Fluorescence, Quantitative RT-PCR

    Characterization of endogenous pluripotent makers in selected iPS cell lines. Panel A. Total RNA was isolated using RNeasy Micro Kit from selected iPS cell lines (G1–G3, G5, G6), hES H9 cells (H9), and human primary fibroblasts (F). Total RNA (500 ng) was reverse-transcribed using Superscript III Reverse Transcriptase primed with oligo(dT) 12–18 and used as template in subsequent PCR with Taq DNA Polymerase. PCR analysis examined the expression of endogenous Oct4, Nanog, Sox2, as well as ABCG2, Rex1, DNMT3B and hTERT. GAPDH was used as an internal control. N, no template control (N). PCR products were analyzed on a 10% polyacrylamide TBE Precast Gel. Panel B. TRAP assay for telomerase activity. Selected iPS cells (G1–G3, G6), hES H9 cells (H9), and human primary fibroblasts (F) were analyzed for telomerase activity using the TRAPEZE RT Telomerase Detection Kit as described in M M. PCR products were separated on 10% polyacrylamide TBE Precast Gel. Individual samples are as indicated.

    Journal: PLoS ONE

    Article Title: Antibody-Directed Lentiviral Gene Transduction for Live-Cell Monitoring and Selection of Human iPS and hES Cells

    doi: 10.1371/journal.pone.0034778

    Figure Lengend Snippet: Characterization of endogenous pluripotent makers in selected iPS cell lines. Panel A. Total RNA was isolated using RNeasy Micro Kit from selected iPS cell lines (G1–G3, G5, G6), hES H9 cells (H9), and human primary fibroblasts (F). Total RNA (500 ng) was reverse-transcribed using Superscript III Reverse Transcriptase primed with oligo(dT) 12–18 and used as template in subsequent PCR with Taq DNA Polymerase. PCR analysis examined the expression of endogenous Oct4, Nanog, Sox2, as well as ABCG2, Rex1, DNMT3B and hTERT. GAPDH was used as an internal control. N, no template control (N). PCR products were analyzed on a 10% polyacrylamide TBE Precast Gel. Panel B. TRAP assay for telomerase activity. Selected iPS cells (G1–G3, G6), hES H9 cells (H9), and human primary fibroblasts (F) were analyzed for telomerase activity using the TRAPEZE RT Telomerase Detection Kit as described in M M. PCR products were separated on 10% polyacrylamide TBE Precast Gel. Individual samples are as indicated.

    Article Snippet: RT-PCR and PCR Total RNA was harvested using RNeasy Micro Kit (Qiagen) and quantified by spectrophotometer.

    Techniques: Isolation, Polymerase Chain Reaction, Expressing, TRAP Assay, Activity Assay

    Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Article Snippet: RNA isolation and quality control RNA from sorted cells was isolated with the use of the RNeasy plus micro kit (Qiagen, 74,034) or the RNAqueous-Micro Total RNA Isolation Kit (Ambion, AM1931).

    Techniques: Lysis, Isolation

    Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Article Snippet: RNA isolation and quality control RNA from sorted cells was isolated with the use of the RNeasy plus micro kit (Qiagen, 74,034) or the RNAqueous-Micro Total RNA Isolation Kit (Ambion, AM1931).

    Techniques: Lysis, Isolation, FACS

    Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Article Snippet: RNA isolation and quality control RNA from sorted cells was isolated with the use of the RNeasy plus micro kit (Qiagen, 74,034) or the RNAqueous-Micro Total RNA Isolation Kit (Ambion, AM1931).

    Techniques: Purification, FACS, Quantitative RT-PCR, Isolation

    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Journal: PLoS ONE

    Article Title: Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein

    doi: 10.1371/journal.pone.0048555

    Figure Lengend Snippet: RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Article Snippet: Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Amplification, Expressing, Microarray, Derivative Assay

    Image showing nucleotide size distribution from CTAB (panel A) and CTAB + RNeasy RNA isolations (panel B) . Nucleotides have been resolved by denaturing gel electrophoresis and have been stained using ethidium bromide and viewed under a UV transilluminator. In both samples, clear rRNA bands can be seen, suggesting the integrity of the RNAs. In the CTAB gel, a band of high-molecular weight DNA can be seen, this is absent from the CTAB + RNeasy extractions.

    Journal: BMC Research Notes

    Article Title: Optimization and comparison of different methods for RNA isolation for cDNA library construction from the reindeer lichen Cladonia rangiferina

    doi: 10.1186/1756-0500-2-204

    Figure Lengend Snippet: Image showing nucleotide size distribution from CTAB (panel A) and CTAB + RNeasy RNA isolations (panel B) . Nucleotides have been resolved by denaturing gel electrophoresis and have been stained using ethidium bromide and viewed under a UV transilluminator. In both samples, clear rRNA bands can be seen, suggesting the integrity of the RNAs. In the CTAB gel, a band of high-molecular weight DNA can be seen, this is absent from the CTAB + RNeasy extractions.

    Article Snippet: The extracted total RNA was cleaned using the RNeasy Midi Kit (Qiagen, Germany) according to the manufacturer's instructions.

    Techniques: Nucleic Acid Electrophoresis, Staining, Molecular Weight

    Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Lysis, Isolation

    Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Lysis, Isolation, FACS

    Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Purification, FACS, Quantitative RT-PCR, Isolation

    Bland-Altman plots for investigating the level of agreement between RNA extraction kits. Each plot shows the differences between the two kits against the averages of the two kits. The lines represent the mean differences and upper and lower limits of agreement (LOA, mean differences ±1.96SD). a Comparison of RNA yield (ng/μl) of samples extracted with High Pure FFPE RNA Micro Kit and RNeasy® FFPE kit. b Comparison of purity (A260/A280) of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. c Comparison of RIN-values of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. d Comparison of RNA yield (ng/μl) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. e Comparison of purity (A260/A280) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. f Comparison of RIN-values of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit

    Journal: BMC Medical Research Methodology

    Article Title: Quantity and quality of nucleic acids extracted from archival formalin fixed paraffin embedded prostate biopsies

    doi: 10.1186/s12874-018-0628-1

    Figure Lengend Snippet: Bland-Altman plots for investigating the level of agreement between RNA extraction kits. Each plot shows the differences between the two kits against the averages of the two kits. The lines represent the mean differences and upper and lower limits of agreement (LOA, mean differences ±1.96SD). a Comparison of RNA yield (ng/μl) of samples extracted with High Pure FFPE RNA Micro Kit and RNeasy® FFPE kit. b Comparison of purity (A260/A280) of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. c Comparison of RIN-values of samples extracted with High Pure FFPE RNA Micro kit and RNeasy® FFPE kit. d Comparison of RNA yield (ng/μl) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. e Comparison of purity (A260/A280) of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit. f Comparison of RIN-values of samples extracted with RNeasy® FFPE kit and AllPrep® DNA/RNA FFPE kit

    Article Snippet: The macro-dissected tumor area from one tissue section was used for RNA isolation using the RNeasy® FFPE kit (Qiagen, Hilden, Germany) and the tumor area from the second section was used for DNA isolation using the QIAamp® DNA FFPE Tissue kit (Qiagen, Hilden, Germany), following the manufacturers’ instructions.

    Techniques: RNA Extraction, Formalin-fixed Paraffin-Embedded

    The decreased levels of CCR5 expression by CCR5 shRNA (1005) correlates with that of CCR5 mRNA . huCCR5-293T cells were transduced with lentiviral vectors bearing either shRNA 1005 against CCR5 {CCR5shRNA (1005)} or a control shRNA against firefly luciferase (Luc shRNA). To monitor the expression levels of CCR5 on cell surface, the cells were cultured for 4 days and stained with either PE-Cy5 conjugated anti human CCR5 monoclonal antibody or isotype control. CCR5 and EGFP expression were analyzed by flow cytometry. The percentage number in each quadrant is indicated in each panel (A). To measure the levels of CCR5 mRNA, total RNA was isolated using Qiagen RNeasy extraction kit. Quantitative RT-PCR was performed using IQ5 with iScript one step RT-PCR kit using β-actin as an internal control (B).

    Journal: Genetic Vaccines and Therapy

    Article Title: Characterization of a potent non-cytotoxic shRNA directed to the HIV-1 co-receptor CCR5

    doi: 10.1186/1479-0556-7-8

    Figure Lengend Snippet: The decreased levels of CCR5 expression by CCR5 shRNA (1005) correlates with that of CCR5 mRNA . huCCR5-293T cells were transduced with lentiviral vectors bearing either shRNA 1005 against CCR5 {CCR5shRNA (1005)} or a control shRNA against firefly luciferase (Luc shRNA). To monitor the expression levels of CCR5 on cell surface, the cells were cultured for 4 days and stained with either PE-Cy5 conjugated anti human CCR5 monoclonal antibody or isotype control. CCR5 and EGFP expression were analyzed by flow cytometry. The percentage number in each quadrant is indicated in each panel (A). To measure the levels of CCR5 mRNA, total RNA was isolated using Qiagen RNeasy extraction kit. Quantitative RT-PCR was performed using IQ5 with iScript one step RT-PCR kit using β-actin as an internal control (B).

    Article Snippet: Quantitative RT-PCR for CCR5 mRNA Total RNAs from the infected CEM NKR-CCR5 cells were isolated using the Qiagen RNeasy extraction kit following manufacture's instruction.

    Techniques: Expressing, shRNA, Transduction, Luciferase, Cell Culture, Staining, Flow Cytometry, Cytometry, Isolation, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction