dynabeads oligo (Thermo Fisher)
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Name:
Dynabeads Oligo(dT)25
Description:
Dynabeads Oligo(dT)25 mRNA isolation beads specifically target and capture mRNA molecules from virtually any crude sample and eliminate the need to purify total RNA when the desired information-bearing nucleic acid is mRNA. Since mRNA comprises only about 1–5% ot total cellular RNA, the isolation of total RNA is not the most efficient way to isolate mRNA. Other technologies designed to purify total RNA yield ~80% ribosomal RNA and force mRNA to compete with ribosomal RNA, transfer RNA, micro RNA, small nucleolar RNA, and small cytoplasmic RNA for membrane binding. Advantages of Dynabeads Oligo(dT)25 beads:• Fast and gentle procedure yields pure intact mRNA• Extremely pure mRNA isolation, best choice upstream of cDNA synthesis• Exquisitely sensitive mRNA isolation enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)How the beads workThe oligo(dT)25-coated Dynabeads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured. Isolated mRNA is pure, eliminating the need for ribosomal RNA subtraction or a post-extraction DNase treatment. This column-free system ensures the highest transcriptome recovery:• Physical mRNA capture on mobile magnetic beads• Rapid and gentle magnetic handling procedures• No mRNA lost during high g-force spins• No mRNA trapped in column membranes during elutionApplicationsmRNA is suitable for all downstream molecular applications, including gene cloning, cDNA synthesis, cDNA library construction, RT-PCR, quantitative RT-PCR, RPA (Ribonuclease Protection Assay), subtractive hybridization, primer extension, SAGE, RACE, and others. The Dynabeads Oligo(dT)25 mRNA isolation beads are the ideal mRNA purification method prior to cDNA library construction. Use of these beads ensures the highest recovery and enrichment of the transcriptome. These beads capture more of the transcriptome than is possible with methods that integrate a total RNA isolation step upstream of mRNA isolation.Verastile elution optionsElution can be performed in any volume down to 5 µL. mRNA elution is optional because enzymatic reactions in downstream procedures are not inhibited by presence of Dynabeads. Additionally, one can perform cDNA synthesis directly on the beads to create a reusable solid-phase cDNA library.
Catalog Number:
61002
Price:
None
Applications:
Automated mRNA Isolation|Bead-Based IVD Assay Development|Bead-Based Nucleic Acid IVD|Clinical|DNA & RNA Purification & Analysis|Diagnostic Development|Molecular Diagnostic Test Development|RNA Extraction|mRNA Isolation|Automated Nucleic Acid Purification
Size:
2 mL
Category:
Beads & Microspheres, Magnetic Beads Ligand-Coupled, Magnetic Beads Oligo(dT)
Score:
85
|
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Structured Review
Thermo Fisher
dynabeads oligo
Dynabeads Oligo(dT)25 mRNA isolation beads specifically target and capture mRNA molecules from virtually any crude sample and eliminate the need to purify total RNA when the desired information-bearing nucleic acid is mRNA. Since mRNA comprises only about 1–5% ot total cellular RNA, the isolation of total RNA is not the most efficient way to isolate mRNA. Other technologies designed to purify total RNA yield ~80% ribosomal RNA and force mRNA to compete with ribosomal RNA, transfer RNA, micro RNA, small nucleolar RNA, and small cytoplasmic RNA for membrane binding. Advantages of Dynabeads Oligo(dT)25 beads:• Fast and gentle procedure yields pure intact mRNA• Extremely pure mRNA isolation, best choice upstream of cDNA synthesis• Exquisitely sensitive mRNA isolation enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)How the beads workThe oligo(dT)25-coated Dynabeads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured. Isolated mRNA is pure, eliminating the need for ribosomal RNA subtraction or a post-extraction DNase treatment. This column-free system ensures the highest transcriptome recovery:• Physical mRNA capture on mobile magnetic beads• Rapid and gentle magnetic handling procedures• No mRNA lost during high g-force spins• No mRNA trapped in column membranes during elutionApplicationsmRNA is suitable for all downstream molecular applications, including gene cloning, cDNA synthesis, cDNA library construction, RT-PCR, quantitative RT-PCR, RPA (Ribonuclease Protection Assay), subtractive hybridization, primer extension, SAGE, RACE, and others. The Dynabeads Oligo(dT)25 mRNA isolation beads are the ideal mRNA purification method prior to cDNA library construction. Use of these beads ensures the highest recovery and enrichment of the transcriptome. These beads capture more of the transcriptome than is possible with methods that integrate a total RNA isolation step upstream of mRNA isolation.Verastile elution optionsElution can be performed in any volume down to 5 µL. mRNA elution is optional because enzymatic reactions in downstream procedures are not inhibited by presence of Dynabeads. Additionally, one can perform cDNA synthesis directly on the beads to create a reusable solid-phase cDNA library.
https://www.bioz.com/result/dynabeads oligo/product/Thermo Fisher
Average 99 stars, based on 113 article reviews
Price from $9.99 to $1999.99
Dynabeads Oligo(dT)25 mRNA isolation beads specifically target and capture mRNA molecules from virtually any crude sample and eliminate the need to purify total RNA when the desired information-bearing nucleic acid is mRNA. Since mRNA comprises only about 1–5% ot total cellular RNA, the isolation of total RNA is not the most efficient way to isolate mRNA. Other technologies designed to purify total RNA yield ~80% ribosomal RNA and force mRNA to compete with ribosomal RNA, transfer RNA, micro RNA, small nucleolar RNA, and small cytoplasmic RNA for membrane binding. Advantages of Dynabeads Oligo(dT)25 beads:• Fast and gentle procedure yields pure intact mRNA• Extremely pure mRNA isolation, best choice upstream of cDNA synthesis• Exquisitely sensitive mRNA isolation enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)How the beads workThe oligo(dT)25-coated Dynabeads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured. Isolated mRNA is pure, eliminating the need for ribosomal RNA subtraction or a post-extraction DNase treatment. This column-free system ensures the highest transcriptome recovery:• Physical mRNA capture on mobile magnetic beads• Rapid and gentle magnetic handling procedures• No mRNA lost during high g-force spins• No mRNA trapped in column membranes during elutionApplicationsmRNA is suitable for all downstream molecular applications, including gene cloning, cDNA synthesis, cDNA library construction, RT-PCR, quantitative RT-PCR, RPA (Ribonuclease Protection Assay), subtractive hybridization, primer extension, SAGE, RACE, and others. The Dynabeads Oligo(dT)25 mRNA isolation beads are the ideal mRNA purification method prior to cDNA library construction. Use of these beads ensures the highest recovery and enrichment of the transcriptome. These beads capture more of the transcriptome than is possible with methods that integrate a total RNA isolation step upstream of mRNA isolation.Verastile elution optionsElution can be performed in any volume down to 5 µL. mRNA elution is optional because enzymatic reactions in downstream procedures are not inhibited by presence of Dynabeads. Additionally, one can perform cDNA synthesis directly on the beads to create a reusable solid-phase cDNA library.
https://www.bioz.com/result/dynabeads oligo/product/Thermo Fisher
Average 99 stars, based on 113 article reviews
Price from $9.99 to $1999.99
dynabeads oligo - by Bioz Stars,
2019-12
99/100 stars
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Amplification:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Fragments of 200-400 bp were size-selected using Agencourt AMPure XP beads, and subjected to PCR amplification using Q5 DNA polymerase (New England Biolabs). .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: In short, mRNA was isolated from 1 μg total RNA using Article Title: Endogenous retroviruses function as species-specific enhancer elements in the placenta Article Snippet: Two replicates from different passages (10 million cells each) were prepared for each cell type. mRNA was extracted directly from cell lysates using Expressing:Article Title: Late-stage tumors induce anemia and immunosuppressive extramedullary erythroid progenitor cells Article Snippet: The RNAseq library for these RNA samples was constructed according to the strand-specific RNA sequencing library preparation protocol. mRNA transcripts were enriched by two rounds of poly-(A+) selection with Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: Paragraph title: Expression Studies ... For Northern blots mRNA was isolated from 75 μg of total RNA using Article Title: Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma Article Snippet: 3′ RNA-SAGE (serial analysis of gene expression) sequencing was performed on ABI SOLiD platform by Mission Biotech (Taiwan) according to manufacturer’s protocol and 10–23 million reads were generated from each sampler ( ). .. Briefly, mRNA was purified using Article Title: Brown adipose tissue macrophages control tissue innervation and homeostatic energy expenditure Article Snippet: In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment Article Snippet: The original scan of western blots are provided as . .. RNAs isolated from MDA231-LM2 cells expressing shCntr or shGALNT14 and MDA231-Par cells expressing pBabe-Hygro vector or GALNT14 were subjected to enrichment of polyA+ RNA using Article Title: Genome-wide association study implicates NDST3 in schizophrenia and bipolar disorder Article Snippet: Paragraph title: Genome-wide expression profiling using RNA-seq technology ... Total RNA was extracted from ~100 mg of hippocampus tissue and mRNA was isolated from 35 μg of total RNA with Synthesized:Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: For Northern blots mRNA was isolated from 75 μg of total RNA using Construct:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Barcodes were utilized for multiplexing. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Late-stage tumors induce anemia and immunosuppressive extramedullary erythroid progenitor cells Article Snippet: Total RNA from sorted CD45+ CD71+ TER119+ and CD45-CD71+ TER119+ cells from neonatal, anemic and tumor-bearing (LLC) mice and MDSCs from tumor-bearing (LLC) mice was extracted using Trizol reagent (Life Technologies) and then treated with DNase I (Qiagen). .. The RNAseq library for these RNA samples was constructed according to the strand-specific RNA sequencing library preparation protocol. mRNA transcripts were enriched by two rounds of poly-(A+) selection with SYBR Green Assay:Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: For Northern blots mRNA was isolated from 75 μg of total RNA using Incubation:Article Title: The RNA helicase UPF1 associates with mRNAs co-transcriptionally and is required for the release of mRNAs from gene loci Article Snippet: The TRI Reagent–pellet suspension was incubated at 65°C with intermittent vortexing to dissolve the pellet, and RNA was extracted following the manufacturer’s protocol. .. Poly(A) depletion was performed with Article Title: Novel Method for High-Throughput Full-Length IGHV-D-J Sequencing of the Immune Repertoire from Bulk B-Cells with Single-Cell Resolution Article Snippet: PBMCs from normal blood donors were incubated with the following anti-human Abs: V500 anti-CD19 (BD Biosciences), PerCPcy5.5 anti-CD38 (BioLegend), PE-cy7 anti- CD24 (BioLegend), FITC anti-IgD (ThermoFisher), and allophycocyanin anti-CD27 (BD Bioscien). .. B cells were sorted directly into 200 µl PCR tubes containing 100 µl Mass Spectrometry:Article Title: Thiol-linked alkylation of RNA to assess expression dynamics Article Snippet: Data were interpreted using the Trace Finder software suite (Thermo Fisher Scientific) and manually validated. .. To determine s4 U incorporation events in polyadenylated or total RNA by Mass Spectrometry, total RNA was either subjected to oligo(dT) enrichment using Genome Wide:Article Title: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment Article Snippet: Paragraph title: Genome-wide RNA-seq analysis ... RNAs isolated from MDA231-LM2 cells expressing shCntr or shGALNT14 and MDA231-Par cells expressing pBabe-Hygro vector or GALNT14 were subjected to enrichment of polyA+ RNA using Article Title: Genome-wide association study implicates NDST3 in schizophrenia and bipolar disorder Article Snippet: Paragraph title: Genome-wide expression profiling using RNA-seq technology ... Total RNA was extracted from ~100 mg of hippocampus tissue and mRNA was isolated from 35 μg of total RNA with Derivative Assay:Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: In short, mRNA was isolated from 1 μg total RNA using High Performance Liquid Chromatography:Article Title: Thiol-linked alkylation of RNA to assess expression dynamics Article Snippet: Acidified samples were separated on a Ultimate U300 BioRSLC HPLC system (Dionex; Thermo Fisher Scientific), employing a Kinetex F5 Pentafluorophenyl column (150 mm x 2.1 mm; 2.6 µm, 100 Å; Phenomenex) with a flow rate of 100 µl/min. .. To determine s4 U incorporation events in polyadenylated or total RNA by Mass Spectrometry, total RNA was either subjected to oligo(dT) enrichment using Flow Cytometry:Article Title: Thiol-linked alkylation of RNA to assess expression dynamics Article Snippet: Acidified samples were separated on a Ultimate U300 BioRSLC HPLC system (Dionex; Thermo Fisher Scientific), employing a Kinetex F5 Pentafluorophenyl column (150 mm x 2.1 mm; 2.6 µm, 100 Å; Phenomenex) with a flow rate of 100 µl/min. .. To determine s4 U incorporation events in polyadenylated or total RNA by Mass Spectrometry, total RNA was either subjected to oligo(dT) enrichment using Ligation:Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: In short, mRNA was isolated from 1 μg total RNA using Protease Inhibitor:Article Title: The RNA helicase UPF1 associates with mRNAs co-transcriptionally and is required for the release of mRNAs from gene loci Article Snippet: Equal volume of 2X NUN buffer (50 mM HEPES-KOH pH 7.6, 600 mM NaCl, 2 M Urea, 2% NP-40, 1 mM DTT, 1X cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail and 1 U/µL Ribolock RNase Inhibitor) was added to the nuclear suspension drop by drop while vortexing and the suspension was placed on ice for 20 min prior to spinning at 13,000 RPM for 30 min at 4°C. .. Poly(A) depletion was performed with Northern Blot:Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: RNAs were extracted with Trizol (Invitrogen), following manufacturer’s guidelines. .. For Northern blots mRNA was isolated from 75 μg of total RNA using Cell Culture:Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Briefly, total RNA was extracted from cultured cells using the RNeasy Plus Mini Kit (QIAGEN). .. Poly(A) RNA was isolated using Generated:Article Title: Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma Article Snippet: 3′ RNA-SAGE (serial analysis of gene expression) sequencing was performed on ABI SOLiD platform by Mission Biotech (Taiwan) according to manufacturer’s protocol and 10–23 million reads were generated from each sampler ( ). .. Briefly, mRNA was purified using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: In short, mRNA was isolated from 1 μg total RNA using DNA Sequencing:Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Polymerase Chain Reaction:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Fragments of 200-400 bp were size-selected using Agencourt AMPure XP beads, and subjected to PCR amplification using Q5 DNA polymerase (New England Biolabs). .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: Endogenous retroviruses function as species-specific enhancer elements in the placenta Article Snippet: Two replicates from different passages (10 million cells each) were prepared for each cell type. mRNA was extracted directly from cell lysates using Article Title: Novel Method for High-Throughput Full-Length IGHV-D-J Sequencing of the Immune Repertoire from Bulk B-Cells with Single-Cell Resolution Article Snippet: Non-B cells were excluded with efluor-450 anti-CD3 and anti-CD16, and dead cells were excluded by Sytox Blue staining (ThermoFisher). .. B cells were sorted directly into 200 µl PCR tubes containing 100 µl Binding Assay:Article Title: The spliceosome is a therapeutic vulnerability in MYC-driven cancer Article Snippet: Isolation of newly transcribed RNA was performed as described using 100 µL streptavidin beads (Miltenyi Biotec). .. Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: For Northern blots mRNA was isolated from 75 μg of total RNA using ChIP-sequencing:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: For RNA-seq libraries, polyA+ RNA was isolated using Dynabeads Oligo(dT)25 (Invitrogen) and constructed into strand-specific libraries using the dUTP method. .. For RNA-seq libraries, polyA+ RNA was isolated using Multiplexing:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Barcodes were utilized for multiplexing. .. For RNA-seq libraries, polyA+ RNA was isolated using RNA Sequencing Assay:Article Title: Global translational reprogramming is a fundamental layer of immune regulation in plants Article Snippet: Paragraph title: RNA-seq library construction ... 50–75 μg total RNA was used for mRNA purification with Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Barcodes were utilized for multiplexing. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Late-stage tumors induce anemia and immunosuppressive extramedullary erythroid progenitor cells Article Snippet: Total RNA from sorted CD45+ CD71+ TER119+ and CD45-CD71+ TER119+ cells from neonatal, anemic and tumor-bearing (LLC) mice and MDSCs from tumor-bearing (LLC) mice was extracted using Trizol reagent (Life Technologies) and then treated with DNase I (Qiagen). .. The RNAseq library for these RNA samples was constructed according to the strand-specific RNA sequencing library preparation protocol. mRNA transcripts were enriched by two rounds of poly-(A+) selection with Article Title: Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma Article Snippet: Briefly, mRNA was purified using Article Title: Re-evaluating Microglia Expression Profiles Using RiboTag and Cell Isolation Strategies Article Snippet: Paragraph title: RNA sequencing ... In brief, 5,000 microglia cells were sorted into 50μl of Lysis Buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: Cell lines were tested for mycoplasma contamination on a monthly basis. .. For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: Paragraph title: RNA-sequencing (RNA-seq) and data analysis ... In short, mRNA was isolated from 1 μg total RNA using Article Title: Endogenous retroviruses function as species-specific enhancer elements in the placenta Article Snippet: Paragraph title: 3′ RNA-Seq ... Two replicates from different passages (10 million cells each) were prepared for each cell type. mRNA was extracted directly from cell lysates using Article Title: Brown adipose tissue macrophages control tissue innervation and homeostatic energy expenditure Article Snippet: In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Dynabeads oligo(dT) (Life Technologies) according to manufacturer’s guidelines. .. In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Poly(A) RNA was isolated using Dynabeads Oligo(dT)25 (Invitrogen) and used as input for library construction utilizing the dUTP method as described50 . .. Poly(A) RNA was isolated using Article Title: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment Article Snippet: Paragraph title: Genome-wide RNA-seq analysis ... RNAs isolated from MDA231-LM2 cells expressing shCntr or shGALNT14 and MDA231-Par cells expressing pBabe-Hygro vector or GALNT14 were subjected to enrichment of polyA+ RNA using Article Title: Genome-Wide Profiling of Cervical RNA-Binding Proteins Identifies Human Papillomavirus Regulation of RNASEH2A Expression by Viral E7 and E2F1 Article Snippet: RNA-seq 1 was performed as described previously ( , ). .. Briefly, polyadenylated [poly(A)+ ] RNA was obtained with two rounds of poly(A) selection using Article Title: Genome-wide association study implicates NDST3 in schizophrenia and bipolar disorder Article Snippet: Paragraph title: Genome-wide expression profiling using RNA-seq technology ... Total RNA was extracted from ~100 mg of hippocampus tissue and mRNA was isolated from 35 μg of total RNA with Fluorescence:Article Title: The spliceosome is a therapeutic vulnerability in MYC-driven cancer Article Snippet: Isolation:Article Title: The spliceosome is a therapeutic vulnerability in MYC-driven cancer Article Snippet: Paragraph title: Poly(A)+ RNA isolation ... Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Barcodes were utilized for multiplexing. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: RNAs were extracted with Trizol (Invitrogen), following manufacturer’s guidelines. .. For Northern blots mRNA was isolated from 75 μg of total RNA using Article Title: Novel mechanisms for organic acid-mediated aluminium tolerance in roots and leaves of two contrasting soybean genotypes Article Snippet: Total RNA was isolated by using Trizol reagent (Takara, Dalian, China). .. The mRNAs were purified by using Article Title: Defective transcription elongation in a subset of cancers confers immunotherapy resistance Article Snippet: Total RNA was extracted from the cells using Tri reagent (Sigma), followed by rRNA depletion and subsequent concentration of rRNA-depleted samples using RiboMinus™ Eukaryote Kit (Ambion) according to manufacturer’s instructions. .. PolyA + -RNA was isolated from rRNA-depleted samples using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: RNA-seq libraries were prepared essentially as described ( ). .. In short, mRNA was isolated from 1 μg total RNA using Article Title: Brown adipose tissue macrophages control tissue innervation and homeostatic energy expenditure Article Snippet: Paragraph title: RNA Isolation, Library Construction, and Analysis ... In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: The RNA helicase UPF1 associates with mRNAs co-transcriptionally and is required for the release of mRNAs from gene loci Article Snippet: Paragraph title: Nascent RNA isolation from S2 cells ... Poly(A) depletion was performed with Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Briefly, total RNA was extracted from cultured cells using the RNeasy Plus Mini Kit (QIAGEN). .. Poly(A) RNA was isolated using Article Title: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment Article Snippet: The original scan of western blots are provided as . .. RNAs isolated from MDA231-LM2 cells expressing shCntr or shGALNT14 and MDA231-Par cells expressing pBabe-Hygro vector or GALNT14 were subjected to enrichment of polyA+ RNA using Article Title: Genome-Wide Profiling of Cervical RNA-Binding Proteins Identifies Human Papillomavirus Regulation of RNASEH2A Expression by Viral E7 and E2F1 Article Snippet: Paragraph title: RNA isolation and sequencing. ... Briefly, polyadenylated [poly(A)+ ] RNA was obtained with two rounds of poly(A) selection using Article Title: Genome-wide association study implicates NDST3 in schizophrenia and bipolar disorder Article Snippet: Rat study protocols were approved by the Institutional Animal Care and Use Committee at Indiana University School of Medicine. .. Total RNA was extracted from ~100 mg of hippocampus tissue and mRNA was isolated from 35 μg of total RNA with Transfection:Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: Cell lines were tested for mycoplasma contamination on a monthly basis. .. For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Mouse Assay:Article Title: Late-stage tumors induce anemia and immunosuppressive extramedullary erythroid progenitor cells Article Snippet: Total RNA from sorted CD45+ CD71+ TER119+ and CD45-CD71+ TER119+ cells from neonatal, anemic and tumor-bearing (LLC) mice and MDSCs from tumor-bearing (LLC) mice was extracted using Trizol reagent (Life Technologies) and then treated with DNase I (Qiagen). .. The RNAseq library for these RNA samples was constructed according to the strand-specific RNA sequencing library preparation protocol. mRNA transcripts were enriched by two rounds of poly-(A+) selection with Sequencing:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Libraries were quantified by qPCR using primers annealing to the adapter sequence and sequenced at a concentration of 12 pM on an Illumina HiSeq. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma Article Snippet: 3′ RNA-SAGE (serial analysis of gene expression) sequencing was performed on ABI SOLiD platform by Mission Biotech (Taiwan) according to manufacturer’s protocol and 10–23 million reads were generated from each sampler ( ). .. Briefly, mRNA was purified using Article Title: Novel mechanisms for organic acid-mediated aluminium tolerance in roots and leaves of two contrasting soybean genotypes Article Snippet: Paragraph title: RNA isolation, library construction and high-throughput sequencing ... The mRNAs were purified by using Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: Paragraph title: Library preparation and high-throughput sequencing ... For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: JUNB governs a feed-forward network of TGFβ signaling that aggravates breast cancer invasion Article Snippet: In short, mRNA was isolated from 1 μg total RNA using Article Title: Endogenous retroviruses function as species-specific enhancer elements in the placenta Article Snippet: Two replicates from different passages (10 million cells each) were prepared for each cell type. mRNA was extracted directly from cell lysates using Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Poly(A) RNA was isolated using Article Title: Genome-Wide Profiling of Cervical RNA-Binding Proteins Identifies Human Papillomavirus Regulation of RNASEH2A Expression by Viral E7 and E2F1 Article Snippet: Paragraph title: RNA isolation and sequencing. ... Briefly, polyadenylated [poly(A)+ ] RNA was obtained with two rounds of poly(A) selection using Article Title: Genome-wide association study implicates NDST3 in schizophrenia and bipolar disorder Article Snippet: Total RNA was extracted from ~100 mg of hippocampus tissue and mRNA was isolated from 35 μg of total RNA with FACS:Article Title: Novel Method for High-Throughput Full-Length IGHV-D-J Sequencing of the Immune Repertoire from Bulk B-Cells with Single-Cell Resolution Article Snippet: Paragraph title: Cell Sorting ... B cells were sorted directly into 200 µl PCR tubes containing 100 µl Purification:Article Title: Global translational reprogramming is a fundamental layer of immune regulation in plants Article Snippet: 0.75 ml TRIzol® LS (Ambion) was added to the 0.25 ml lysate saved from the Ribo-seq library construction, from which total RNA was extracted, quantified and qualified using Nanodrop (Thermo Fisher Scientific Inc). .. 50–75 μg total RNA was used for mRNA purification with Article Title: Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma Article Snippet: 3′ RNA-SAGE (serial analysis of gene expression) sequencing was performed on ABI SOLiD platform by Mission Biotech (Taiwan) according to manufacturer’s protocol and 10–23 million reads were generated from each sampler ( ). .. Briefly, mRNA was purified using Article Title: Novel mechanisms for organic acid-mediated aluminium tolerance in roots and leaves of two contrasting soybean genotypes Article Snippet: Total RNA was isolated by using Trizol reagent (Takara, Dalian, China). .. The mRNAs were purified by using Article Title: Endogenous retroviruses function as species-specific enhancer elements in the placenta Article Snippet: Two replicates from different passages (10 million cells each) were prepared for each cell type. mRNA was extracted directly from cell lysates using Plasmid Preparation:Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment Article Snippet: The original scan of western blots are provided as . .. RNAs isolated from MDA231-LM2 cells expressing shCntr or shGALNT14 and MDA231-Par cells expressing pBabe-Hygro vector or GALNT14 were subjected to enrichment of polyA+ RNA using Software:Article Title: Thiol-linked alkylation of RNA to assess expression dynamics Article Snippet: Data were interpreted using the Trace Finder software suite (Thermo Fisher Scientific) and manually validated. .. To determine s4 U incorporation events in polyadenylated or total RNA by Mass Spectrometry, total RNA was either subjected to oligo(dT) enrichment using Article Title: Brown adipose tissue macrophages control tissue innervation and homeostatic energy expenditure Article Snippet: In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Real-time Polymerase Chain Reaction:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Libraries were quantified by qPCR using primers annealing to the adapter sequence and sequenced at a concentration of 12 pM on an Illumina HiSeq. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Postnatal loss of Dlk1 imprinting in stem cells and niche-astrocytes regulates neurogenesis Article Snippet: For Northern blots mRNA was isolated from 75 μg of total RNA using RNA Extraction:Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Paragraph title: RNA Extraction and Next-Generation Sequencing Analysis ... Poly(A) RNA was isolated using Selection:Article Title: Late-stage tumors induce anemia and immunosuppressive extramedullary erythroid progenitor cells Article Snippet: Total RNA from sorted CD45+ CD71+ TER119+ and CD45-CD71+ TER119+ cells from neonatal, anemic and tumor-bearing (LLC) mice and MDSCs from tumor-bearing (LLC) mice was extracted using Trizol reagent (Life Technologies) and then treated with DNase I (Qiagen). .. The RNAseq library for these RNA samples was constructed according to the strand-specific RNA sequencing library preparation protocol. mRNA transcripts were enriched by two rounds of poly-(A+) selection with Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: Cell lines were tested for mycoplasma contamination on a monthly basis. .. For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Article Title: Genome-Wide Profiling of Cervical RNA-Binding Proteins Identifies Human Papillomavirus Regulation of RNASEH2A Expression by Viral E7 and E2F1 Article Snippet: RNA-seq 1 was performed as described previously ( , ). .. Briefly, polyadenylated [poly(A)+ ] RNA was obtained with two rounds of poly(A) selection using Sample Prep:Article Title: Genome-Wide Profiling of Cervical RNA-Binding Proteins Identifies Human Papillomavirus Regulation of RNASEH2A Expression by Viral E7 and E2F1 Article Snippet: Briefly, polyadenylated [poly(A)+ ] RNA was obtained with two rounds of poly(A) selection using Next-Generation Sequencing:Article Title: The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in mouse embryonic stem cells Article Snippet: Paragraph title: RNA Extraction and Next-Generation Sequencing Analysis ... Poly(A) RNA was isolated using Concentration Assay:Article Title: CO-REPRESSOR CBFA2T2 REGULATES PLURIPOTENCY AND GERMLINE DEVELOPMENT Article Snippet: Libraries were quantified by qPCR using primers annealing to the adapter sequence and sequenced at a concentration of 12 pM on an Illumina HiSeq. .. For RNA-seq libraries, polyA+ RNA was isolated using Article Title: Novel mechanisms for organic acid-mediated aluminium tolerance in roots and leaves of two contrasting soybean genotypes Article Snippet: The mRNAs were purified by using Article Title: Defective transcription elongation in a subset of cancers confers immunotherapy resistance Article Snippet: Total RNA was extracted from the cells using Tri reagent (Sigma), followed by rRNA depletion and subsequent concentration of rRNA-depleted samples using RiboMinus™ Eukaryote Kit (Ambion) according to manufacturer’s instructions. .. PolyA + -RNA was isolated from rRNA-depleted samples using Article Title: Re-evaluating Microglia Expression Profiles Using RiboTag and Cell Isolation Strategies Article Snippet: In brief, 5,000 microglia cells were sorted into 50μl of Lysis Buffer (Life Technologies) and stored at -80°C. mRNA was captured with Lysis:Article Title: The spliceosome is a therapeutic vulnerability in MYC-driven cancer Article Snippet: Isolation of newly transcribed RNA was performed as described using 100 µL streptavidin beads (Miltenyi Biotec). .. Article Title: Re-evaluating Microglia Expression Profiles Using RiboTag and Cell Isolation Strategies Article Snippet: RNA-Seq of populations was performed as described previously . .. In brief, 5,000 microglia cells were sorted into 50μl of Lysis Buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: Brown adipose tissue macrophages control tissue innervation and homeostatic energy expenditure Article Snippet: RNA-Seq was performed as described previously . .. In brief, 103 –105 cells from each population were sorted into 50μl of lysis/binding buffer (Life Technologies) and stored at -80°C. mRNA was captured with Article Title: The RNA helicase UPF1 associates with mRNAs co-transcriptionally and is required for the release of mRNAs from gene loci Article Snippet: The supernatant was removed and the pellet was resuspended in 5 volumes of nuclear lysis buffer (10 mM HEPES-KOH pH 7.6, 100 mM KCl, 0.1 mM EDTA, 10% Glycerol, 0.15 mM Spermine, 0.5 mM Spermidine, 0.1 M NaF, 0.1 M Na3VO4, 0.1 mM ZnCl2, 1 mM DTT, 1X cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail and 1 U/µL Ribolock RNase Inhibitor) and resuspended using a 2 mL Dounce homogenizer by three strokes with loose pestle and two strokes with tight pestle. .. Poly(A) depletion was performed with Article Title: Novel Method for High-Throughput Full-Length IGHV-D-J Sequencing of the Immune Repertoire from Bulk B-Cells with Single-Cell Resolution Article Snippet: Non-B cells were excluded with efluor-450 anti-CD3 and anti-CD16, and dead cells were excluded by Sytox Blue staining (ThermoFisher). .. B cells were sorted directly into 200 µl PCR tubes containing 100 µl High Throughput Screening Assay:Article Title: Novel mechanisms for organic acid-mediated aluminium tolerance in roots and leaves of two contrasting soybean genotypes Article Snippet: Paragraph title: RNA isolation, library construction and high-throughput sequencing ... The mRNAs were purified by using Article Title: Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis Article Snippet: Cell lines were tested for mycoplasma contamination on a monthly basis. .. For preparation of high-throughput RNA sequencing (RNA-seq) libraries, the total RNA obtained from transfected HEK293T cells or MCF7 cells was enriched for mRNA by performing polyA selection of 20 µg of total RNA using Staining:Article Title: Novel Method for High-Throughput Full-Length IGHV-D-J Sequencing of the Immune Repertoire from Bulk B-Cells with Single-Cell Resolution Article Snippet: Non-B cells were excluded with efluor-450 anti-CD3 and anti-CD16, and dead cells were excluded by Sytox Blue staining (ThermoFisher). .. B cells were sorted directly into 200 µl PCR tubes containing 100 µl |