multiplex small rna library prep kit  (New England Biolabs)


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    Name:
    NEBNext Ultra RNA Library Prep Kit for Illumina
    Description:

    Catalog Number:
    E7530L
    Price:
    None
    Score:
    85
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    Structured Review

    New England Biolabs multiplex small rna library prep kit
    Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from <t>immunoprecipitated</t> MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small <t>RNA</t> libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries

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    Images

    1) Product Images from "PNLDC1 is essential for piRNA 3′ end trimming and transposon silencing during spermatogenesis in mice"

    Article Title: PNLDC1 is essential for piRNA 3′ end trimming and transposon silencing during spermatogenesis in mice

    Journal: Nature Communications

    doi: 10.1038/s41467-017-00854-4

    Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small RNA libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries
    Figure Legend Snippet: Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small RNA libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries

    Techniques Used: Mouse Assay, Labeling, Autoradiography, Isolation, Immunoprecipitation, Western Blot, Molecular Weight, Marker

    2) Product Images from "Mitochondrial membrane-based initial separation of MIWI and MILI functions during pachytene piRNA biogenesis"

    Article Title: Mitochondrial membrane-based initial separation of MIWI and MILI functions during pachytene piRNA biogenesis

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gky1281

    Increased piRNA lengths and reduced piRNA levels in adult Tdrkh cKO testes. ( A ) Severe reduction of total piRNAs in Tdrkh cKO testes. Total RNA from adult WT and Tdrkh cKO testes were end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. ( B ) The length distribution of small RNAs from adult WT and Tdrkh cKO testicular small RNA libraries. Data were normalized by miRNA reads (21–23nt). ( C ) Extension of MILI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MILI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M: molecular weight marker. ( D ) Diminished MIWI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MIWI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MIWI antibody to show immunoprecipitation efficiency. M: molecular weight marker. ( E ) The length distribution of MILI-piRNAs from adult WT and Tdrkh cKO MILI-piRNA libraries. ( F ) The length distribution of MIWI-piRNAs from adult WT and Tdrkh cKO MIWI-piRNA libraries.
    Figure Legend Snippet: Increased piRNA lengths and reduced piRNA levels in adult Tdrkh cKO testes. ( A ) Severe reduction of total piRNAs in Tdrkh cKO testes. Total RNA from adult WT and Tdrkh cKO testes were end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. ( B ) The length distribution of small RNAs from adult WT and Tdrkh cKO testicular small RNA libraries. Data were normalized by miRNA reads (21–23nt). ( C ) Extension of MILI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MILI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M: molecular weight marker. ( D ) Diminished MIWI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MIWI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MIWI antibody to show immunoprecipitation efficiency. M: molecular weight marker. ( E ) The length distribution of MILI-piRNAs from adult WT and Tdrkh cKO MILI-piRNA libraries. ( F ) The length distribution of MIWI-piRNAs from adult WT and Tdrkh cKO MIWI-piRNA libraries.

    Techniques Used: Labeling, Autoradiography, Isolation, Immunoprecipitation, Western Blot, Molecular Weight, Marker

    Related Articles

    Centrifugation:

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    Amplification:

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    Construct:

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    Electrophoresis:

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    IA:

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    Cell Culture:

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    Article Snippet: Cells transfected with the sgCTRL were cultured in the presence of TNFα (10 ng/ml) or left untreated to serve as a control. .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent.

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
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    Expressing:

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    Derivative Assay:

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    Transfection:

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    Ligation:

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    Dissection:

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    Infection:

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    Generated:

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    Methylation:

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
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    Sequencing:

    Article Title: Abnormal neutrophil signature in the blood and pancreas of presymptomatic and symptomatic type 1 diabetes
    Article Snippet: The cDNA library was prepared using a NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s instructions. .. The libraries were hybridized to the flow cell and cluster was generated by bridge amplification (TruSeq SBS Kit v3-HS, Illumina).

    Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)
    Article Snippet: The total RNAs for each group were homogenized to maintain the same concentrations. .. Sequencing libraries were constructed with NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, Beijing, China). .. Two MC groups were independently sequenced, respectively.

    Article Title: Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells
    Article Snippet: Paragraph title: RNA-Seq library preparation and sequencing ... 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide.

    Article Title: Identification and characterization of NF-Y gene family in walnut (Juglans regia L.)
    Article Snippet: A total of 1.5 μg RNA per sample was used as input material for the RNA sample preparations. .. Sequencing libraries were generated using NEBNext ® Ultra™ RNA Library Prep Kit for Illumina ® (NEB, USA). .. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia).

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations. .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Article Title: Studying on the strictly self-compatibility mechanism of ‘Liuyefeitao’ peach (Prunus persica L.)
    Article Snippet: Paragraph title: RNA library preparation and sequencing ... Transcriptome libraries were constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs) according to the manufacturer’s instructions.

    Article Title: Generation and classification of transcriptomes in two Croomia species and molecular evolution of CYC/TB1 genes in Stemonaceae
    Article Snippet: RNA quality and quantity were assessed using gel electrophoresis and a NanoDrop spectrophotometer 2000 (Thermo Scientific, Wilmington, DE, USA). .. Sequencing libraries were generated from 3 μg total RNA using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's protocol, and index codes were added to attribute sequences to each sample. .. After validating the library quality with an Agilent 2100 Bioanalyzer system, the constructed libraries were sequenced on an Illumina Hiseq 2000 platform and paired-end reads were generated.

    Article Title: Functional Insights into the Roles of Hormones in the Dendrobium officinale-Tulasnella sp. Germinated Seed Symbiotic Association
    Article Snippet: For each sample, 3 µg of the total RNA was used as input material for the RNA-Seq library preparations. .. Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswitch, MA, USA) following the manufacturer’s recommendations, and index codes were added to attribute the sequencing reads to each sample. .. The PCR products were purified (AMPure XP system) and the library quality was assessed on the Agilent Bioanalyzer 2100.

    Article Title: Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek’s disease virus
    Article Snippet: A total of 1 μg purified RNA per sample was used for next-generation sequencing (NGS). .. Sequencing libraries were generated using NEBNext®Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer’s instruction, and index codes were added to attribute sequences to each sample as described previously [ ]. .. The libraries were sequenced by Illumina HiSeq 2500.

    Article Title: Comparative transcriptomics identifies genes differentially expressed in the intestine of a new fast-growing strain of common carp with higher unsaturated fatty acid content in muscle
    Article Snippet: Paragraph title: RNA isolation, cDNA library construction and Illumina sequencing ... RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations.

    Article Title: Transcriptome-Wide Analysis of Hepatitis B Virus-Mediated Changes to Normal Hepatocyte Gene Expression
    Article Snippet: The PRHs in the secondary dataset were infected with either AdGFP or AdGFP-HBV 24hr after plating. .. Total RNA was divided for use in downstream miRNA analyses (see below) or for quality control, polyA selection, cDNA library preparation, and the first round of sequencing by Genewiz, Inc (South Plainfield, NJ). cDNA library preparation was done using the NEBNext Ultra RNA Library Preparation Kit (New England Biolabs, Ipswich, MA), and the first round of sequencing was done using the Illumina HiSeq2500 platform to generate 1 x 50bp reads. .. The cDNA library was then used for a second round of sequencing by the DUCOM CGS using the Illumina NextSeq 500 platform to generate an additional set of 1x75bp reads.

    Article Title: Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex
    Article Snippet: Total RNA was isolated from ~25,000–30,000 sorted cells using the Promega Maxwell 16 LEV Simply RNA Cell Kit (Promega). .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent. .. Sequencing was performed on the Illumina HiSeq2500 platform, in a 1 × 50 bp single-read configuration in Rapid Run mode, with a total of at least 120 million reads per lane.

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: Paragraph title: Library preparation and sequencing ... The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art.

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Sequencing experiments for three AT-MSC samples and two hPSC samples and the corresponding pure media were conducted at Exiqon Services, Denmark. .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification.

    Article Title: Sea cucumber genome provides insights into saponin biosynthesis and aestivation regulation
    Article Snippet: Total mRNA was extracted from each sample ( > 1000 embryos/larvae per developmental stage) by following the protocol described by Du et al. . .. All RNA-seq libraries were constructed using the NEB Next mRNA Library Prep Kit by following the manufacturer’s instructions and then were subjected to paired-end 100-bp (PE100) sequencing on the Illumina HiSeq 2000 platform. .. Sequencing reads were aligned to the A . japonicus genome using STAR aligner with its default parameters.

    Nucleic Acid Electrophoresis:

    Article Title: Generation and classification of transcriptomes in two Croomia species and molecular evolution of CYC/TB1 genes in Stemonaceae
    Article Snippet: RNA quality and quantity were assessed using gel electrophoresis and a NanoDrop spectrophotometer 2000 (Thermo Scientific, Wilmington, DE, USA). .. Sequencing libraries were generated from 3 μg total RNA using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's protocol, and index codes were added to attribute sequences to each sample.

    RNA Sequencing Assay:

    Article Title: Abnormal neutrophil signature in the blood and pancreas of presymptomatic and symptomatic type 1 diabetes
    Article Snippet: Paragraph title: Library preparation and RNA-seq ... The cDNA library was prepared using a NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s instructions.

    Article Title: Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells
    Article Snippet: Paragraph title: RNA-Seq library preparation and sequencing ... 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide.

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: RNA with 28S/18S > 1 and an RNA integrity number ≥7 were used for library construction. .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations. .. Libraries were sequenced on the Illumina HiSeq.

    Article Title: Functional Insights into the Roles of Hormones in the Dendrobium officinale-Tulasnella sp. Germinated Seed Symbiotic Association
    Article Snippet: For each sample, 3 µg of the total RNA was used as input material for the RNA-Seq library preparations. .. Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswitch, MA, USA) following the manufacturer’s recommendations, and index codes were added to attribute the sequencing reads to each sample.

    Article Title: Transcriptomic variation of hepatopancreas reveals the energy metabolism and biological processes associated with molting in Chinese mitten crab, Eriocheir sinensis
    Article Snippet: The RNA integrity and quantity were determined using an Agilent 2100 Bioanalyzer (Agilent, Shanghai, China). .. A total of 4 μg RNA with RNA integrity number above 7.0 was used for RNA-Seq library construction using the NEBNext® UltraTM RNA Library Prep Kit for Illumina (NEB, USA). .. Indexed libraries were then pooled and sequenced on Illumina HiseqTM 2500, with 150 bp pair-end reads produced.

    Article Title: Characterization of human plasma-derived exosomal RNAs by deep sequencing
    Article Snippet: To accomplish this goal, we selected plasma samples (samples A, B and C) from three anonymous blood donors and split each sample into two for technical replication. .. We tested the six samples (A1 and A2, B1 and B2, C1 and C2) using two small RNA library preparation kits: NEBNext multiplex small RNA library preparation kit (NEB, New England Biolab, Ipswich, MA, USA) and NEXTflex small RNA sequencing kit (Bioo Scientific, Austin, TX, USA). .. We also tested samples A1 and A2 using the TruSeq small RNA sample preparation kit (Illumina, San Diego, CA, USA).

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Paragraph title: Small RNA sequencing ... NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification.

    Article Title: Global analyses of endonucleolytic cleavage in mammals reveal expanded repertoires of cleavage-inducing small RNAs and their targets
    Article Snippet: Spike-in RNAs (Exiqon) were added into 1 μg total RNA before library construction. .. Small RNA sequencing libraries were generated using NEBNext Small RNA library Prep kit and NEBNext multiplex oligos for Illumina according to the manufacturer's instructions (NEB). .. The final libraries were purified from 6% PAGE gel, and their concentrations were measured using Qubit fluorometric assay (Life Technologies).

    Article Title: Sea cucumber genome provides insights into saponin biosynthesis and aestivation regulation
    Article Snippet: Total mRNA was extracted from each sample ( > 1000 embryos/larvae per developmental stage) by following the protocol described by Du et al. . .. All RNA-seq libraries were constructed using the NEB Next mRNA Library Prep Kit by following the manufacturer’s instructions and then were subjected to paired-end 100-bp (PE100) sequencing on the Illumina HiSeq 2000 platform. .. Sequencing reads were aligned to the A . japonicus genome using STAR aligner with its default parameters.

    Fluorescence:

    Article Title: Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex
    Article Snippet: To control for transfection efficiency, cells were sorted by fluorescence activated cell sorting based on GFP expression using the BD FACSAria II 72 hours posttransfection. .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent.

    Magnetic Beads:

    Article Title: Comparative transcriptomics identifies genes differentially expressed in the intestine of a new fast-growing strain of common carp with higher unsaturated fatty acid content in muscle
    Article Snippet: RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations. .. RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations.

    Isolation:

    Article Title: Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells
    Article Snippet: The RINs of all total RNA samples were 9.7–10. .. 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide. .. Specifically, 100 ng of total RNA together with 2 μL of 1:2000 diluted ERCC was added to extracted mRNA with 15 μL of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94 °C for 8 min, followed by first-strand cDNA synthesis using reverse transcriptase and random primers.

    Article Title: Mating-Induced Transcriptome Changes in the Reproductive Tract of Female Aedes aegypti
    Article Snippet: The concentration of RNA in each sample was measured using a Qubit spectrophotometer (Invitrogen, Grand Island, NY), and the quality of RNA was measured on a Fragment Analyzer (Advanced Analytical Technologies, Inc., Ames, IA). .. From total RNA, mRNA was extracted using the poly-A mRNA Magnetic Isolation Module (New England Biolabs, Inc., Ipswich, MA). cDNA libraries for each replicate were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Inc., Ipswich, MA). .. NEBNext Multiplex Oligos for Illumina (New England Biolabs, Inc., Ipswich, MA) were ligated to each library prior to sequencing.

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: Paragraph title: RNA isolation and RNA-sequencing ... RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Article Title: Comparative transcriptomics identifies genes differentially expressed in the intestine of a new fast-growing strain of common carp with higher unsaturated fatty acid content in muscle
    Article Snippet: Paragraph title: RNA isolation, cDNA library construction and Illumina sequencing ... RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations.

    Article Title: Whole transcriptome profiling reveals the RNA content of motor axons
    Article Snippet: Libraries were pooled and purified with AMPure XP beads for sequencing. .. Three replicate total RNAseq libraries from 5 ng HBRR input each were prepared using the NEBNext Ultra RNA Library Kit for Illumina (NEB) with omission of the mRNA isolation step. .. Instead, fragmentation was performed directly on total RNA by adding 4 μl of first strand reaction buffer (5×) and 1 μl random primers to 5 μl of the 1 ng/μl HBRR/ERCC mix (see above) and heating the mixture at 94°C for 15 min.

    Article Title: Transcriptomic variation of hepatopancreas reveals the energy metabolism and biological processes associated with molting in Chinese mitten crab, Eriocheir sinensis
    Article Snippet: Paragraph title: RNA isolation and RNA-Seq library preparation ... A total of 4 μg RNA with RNA integrity number above 7.0 was used for RNA-Seq library construction using the NEBNext® UltraTM RNA Library Prep Kit for Illumina (NEB, USA).

    Article Title: Transcriptome-Wide Analysis of Hepatitis B Virus-Mediated Changes to Normal Hepatocyte Gene Expression
    Article Snippet: For the secondary dataset, total RNA was isolated using the mirVana RNA isolation kit from triplicate samples of PRHs; these PRHs were isolated from a separate rat liver and were infected with different preparations of AdGFP and AdGFP-HBV than were used to generate the primary dataset. .. Total RNA was divided for use in downstream miRNA analyses (see below) or for quality control, polyA selection, cDNA library preparation, and the first round of sequencing by Genewiz, Inc (South Plainfield, NJ). cDNA library preparation was done using the NEBNext Ultra RNA Library Preparation Kit (New England Biolabs, Ipswich, MA), and the first round of sequencing was done using the Illumina HiSeq2500 platform to generate 1 x 50bp reads.

    Article Title: Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex
    Article Snippet: Total RNA was isolated from ~25,000–30,000 sorted cells using the Promega Maxwell 16 LEV Simply RNA Cell Kit (Promega). .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent.

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art. .. The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art.

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Good performance of Exiqons EV isolation kit has been reported . .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification.

    Flow Cytometry:

    Article Title: Abnormal neutrophil signature in the blood and pancreas of presymptomatic and symptomatic type 1 diabetes
    Article Snippet: The cDNA library was prepared using a NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s instructions. .. The cDNA library was prepared using a NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s instructions.

    Purification:

    Article Title: Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells
    Article Snippet: 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide. .. 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide.

    Article Title: Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek’s disease virus
    Article Snippet: A total of 1 μg purified RNA per sample was used for next-generation sequencing (NGS). .. Sequencing libraries were generated using NEBNext®Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer’s instruction, and index codes were added to attribute sequences to each sample as described previously [ ].

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art. .. The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art.

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Good performance of Exiqons EV isolation kit has been reported . .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification. .. From a total of 50 µl isolated RNA, 6 μl was converted into microRNA NGS libraries.

    Polymerase Chain Reaction:

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art. .. The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art.

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Good performance of Exiqons EV isolation kit has been reported . .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification. .. From a total of 50 µl isolated RNA, 6 μl was converted into microRNA NGS libraries.

    FACS:

    Article Title: Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex
    Article Snippet: To control for transfection efficiency, cells were sorted by fluorescence activated cell sorting based on GFP expression using the BD FACSAria II 72 hours posttransfection. .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent.

    cDNA Library Assay:

    Article Title: Abnormal neutrophil signature in the blood and pancreas of presymptomatic and symptomatic type 1 diabetes
    Article Snippet: Sections were mounted on slides with homemade Mowiol mounting medium (glycerol, G5516; Mowiol 4-88, 81381; and Dabco 33-LV, 290734, Sigma-Aldrich). .. The cDNA library was prepared using a NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s instructions. .. Samples were then purified (AMPure XP Beads, Beckman Coulter) and enriched with PCR to create the final cDNA library.

    Article Title: Generation and classification of transcriptomes in two Croomia species and molecular evolution of CYC/TB1 genes in Stemonaceae
    Article Snippet: Paragraph title: 2.1. Plant samples, cDNA library preparation, and illumina sequencing ... Sequencing libraries were generated from 3 μg total RNA using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's protocol, and index codes were added to attribute sequences to each sample.

    Article Title: Functional Insights into the Roles of Hormones in the Dendrobium officinale-Tulasnella sp. Germinated Seed Symbiotic Association
    Article Snippet: Paragraph title: 4.2. cDNA Library Construction and Nucleotide Sequencing ... Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswitch, MA, USA) following the manufacturer’s recommendations, and index codes were added to attribute the sequencing reads to each sample.

    Article Title: Comparative transcriptomics identifies genes differentially expressed in the intestine of a new fast-growing strain of common carp with higher unsaturated fatty acid content in muscle
    Article Snippet: Paragraph title: RNA isolation, cDNA library construction and Illumina sequencing ... RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations.

    Article Title: Transcriptome-Wide Analysis of Hepatitis B Virus-Mediated Changes to Normal Hepatocyte Gene Expression
    Article Snippet: The PRHs in the secondary dataset were infected with either AdGFP or AdGFP-HBV 24hr after plating. .. Total RNA was divided for use in downstream miRNA analyses (see below) or for quality control, polyA selection, cDNA library preparation, and the first round of sequencing by Genewiz, Inc (South Plainfield, NJ). cDNA library preparation was done using the NEBNext Ultra RNA Library Preparation Kit (New England Biolabs, Ipswich, MA), and the first round of sequencing was done using the Illumina HiSeq2500 platform to generate 1 x 50bp reads. .. The cDNA library was then used for a second round of sequencing by the DUCOM CGS using the Illumina NextSeq 500 platform to generate an additional set of 1x75bp reads.

    RNA Extraction:

    Article Title: Mating-Induced Transcriptome Changes in the Reproductive Tract of Female Aedes aegypti
    Article Snippet: Paragraph title: RNA extraction, library preparation and read processing ... From total RNA, mRNA was extracted using the poly-A mRNA Magnetic Isolation Module (New England Biolabs, Inc., Ipswich, MA). cDNA libraries for each replicate were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Inc., Ipswich, MA).

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: A portion of the mammary biopsies from all IUHT-H and IUCL-H were stored at −80 °C in RNALater until RNA extraction for RNA sequencing. .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Article Title: Generation and classification of transcriptomes in two Croomia species and molecular evolution of CYC/TB1 genes in Stemonaceae
    Article Snippet: Fresh flowers of C. heterosepala (ChFlower) and C. japonica (CjFlower) at the anthesis stage, as well as juvenile leaf samples of C. japonica (CjLeaf), were harvested from these living plants and immediately frozen in liquid nitrogen for storage at −80 °C until total RNA extraction. .. Sequencing libraries were generated from 3 μg total RNA using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's protocol, and index codes were added to attribute sequences to each sample.

    Article Title: Transcriptome-Wide Analysis of Hepatitis B Virus-Mediated Changes to Normal Hepatocyte Gene Expression
    Article Snippet: Paragraph title: RNA extraction, cDNA synthesis, and sequencing ... Total RNA was divided for use in downstream miRNA analyses (see below) or for quality control, polyA selection, cDNA library preparation, and the first round of sequencing by Genewiz, Inc (South Plainfield, NJ). cDNA library preparation was done using the NEBNext Ultra RNA Library Preparation Kit (New England Biolabs, Ipswich, MA), and the first round of sequencing was done using the Illumina HiSeq2500 platform to generate 1 x 50bp reads.

    Plasmid Preparation:

    Article Title: Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex
    Article Snippet: CEM LChIT 3.2 bimodal cells were co-transfected with the plasmid pHAGE EF1α dCas9-VP64 together with sg362F or control sgCTRL and a GFP-expressing plasmid as described above. .. The sequencing library was prepared using the NEBNext Ultra RNA Library Prep Kit reagent.

    Software:

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations. .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Real-time Polymerase Chain Reaction:

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification. .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification.

    Multiplex Assay:

    Article Title: Characterization of human plasma-derived exosomal RNAs by deep sequencing
    Article Snippet: To accomplish this goal, we selected plasma samples (samples A, B and C) from three anonymous blood donors and split each sample into two for technical replication. .. We tested the six samples (A1 and A2, B1 and B2, C1 and C2) using two small RNA library preparation kits: NEBNext multiplex small RNA library preparation kit (NEB, New England Biolab, Ipswich, MA, USA) and NEXTflex small RNA sequencing kit (Bioo Scientific, Austin, TX, USA). .. We also tested samples A1 and A2 using the TruSeq small RNA sample preparation kit (Illumina, San Diego, CA, USA).

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art. .. Poly-A selection with a magnetic isolation module was used to target mRNAs, followed by strand-specific cDNA synthesis with an insert size of 150–400 bp, PCR amplification and library purification.

    Article Title: Global analyses of endonucleolytic cleavage in mammals reveal expanded repertoires of cleavage-inducing small RNAs and their targets
    Article Snippet: Spike-in RNAs (Exiqon) were added into 1 μg total RNA before library construction. .. Small RNA sequencing libraries were generated using NEBNext Small RNA library Prep kit and NEBNext multiplex oligos for Illumina according to the manufacturer's instructions (NEB). .. The final libraries were purified from 6% PAGE gel, and their concentrations were measured using Qubit fluorometric assay (Life Technologies).

    Selection:

    Article Title: Transcriptome-Wide Analysis of Hepatitis B Virus-Mediated Changes to Normal Hepatocyte Gene Expression
    Article Snippet: The PRHs in the secondary dataset were infected with either AdGFP or AdGFP-HBV 24hr after plating. .. Total RNA was divided for use in downstream miRNA analyses (see below) or for quality control, polyA selection, cDNA library preparation, and the first round of sequencing by Genewiz, Inc (South Plainfield, NJ). cDNA library preparation was done using the NEBNext Ultra RNA Library Preparation Kit (New England Biolabs, Ipswich, MA), and the first round of sequencing was done using the Illumina HiSeq2500 platform to generate 1 x 50bp reads. .. The cDNA library was then used for a second round of sequencing by the DUCOM CGS using the Illumina NextSeq 500 platform to generate an additional set of 1x75bp reads.

    Article Title: Whole body transcriptomes and new insights into the biology of the tick Ixodes ricinus
    Article Snippet: The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art. .. The library preparation kit was NEBNext® Ultra Directional RNA Library Prep Kit, NEB Art.

    Ethanol Precipitation:

    Article Title: Mating-Induced Transcriptome Changes in the Reproductive Tract of Female Aedes aegypti
    Article Snippet: RNA was then prepared by chloroform/isopropanol extraction and ethanol precipitation following the manufacturer’s protocol (Life technologies, Grand Island, NY). .. From total RNA, mRNA was extracted using the poly-A mRNA Magnetic Isolation Module (New England Biolabs, Inc., Ipswich, MA). cDNA libraries for each replicate were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Inc., Ipswich, MA).

    Next-Generation Sequencing:

    Article Title: Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek’s disease virus
    Article Snippet: A total of 1 μg purified RNA per sample was used for next-generation sequencing (NGS). .. Sequencing libraries were generated using NEBNext®Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer’s instruction, and index codes were added to attribute sequences to each sample as described previously [ ].

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Good performance of Exiqons EV isolation kit has been reported . .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification. .. From a total of 50 µl isolated RNA, 6 μl was converted into microRNA NGS libraries.

    Spectrophotometry:

    Article Title: Mating-Induced Transcriptome Changes in the Reproductive Tract of Female Aedes aegypti
    Article Snippet: The concentration of RNA in each sample was measured using a Qubit spectrophotometer (Invitrogen, Grand Island, NY), and the quality of RNA was measured on a Fragment Analyzer (Advanced Analytical Technologies, Inc., Ames, IA). .. From total RNA, mRNA was extracted using the poly-A mRNA Magnetic Isolation Module (New England Biolabs, Inc., Ipswich, MA). cDNA libraries for each replicate were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Inc., Ipswich, MA).

    Article Title: Generation and classification of transcriptomes in two Croomia species and molecular evolution of CYC/TB1 genes in Stemonaceae
    Article Snippet: RNA quality and quantity were assessed using gel electrophoresis and a NanoDrop spectrophotometer 2000 (Thermo Scientific, Wilmington, DE, USA). .. Sequencing libraries were generated from 3 μg total RNA using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's protocol, and index codes were added to attribute sequences to each sample.

    Article Title: Functional Insights into the Roles of Hormones in the Dendrobium officinale-Tulasnella sp. Germinated Seed Symbiotic Association
    Article Snippet: RNA purity was checked using the NanoPhotometer® spectrophotometer (Implen, Westlake Vilagge, CA, USA). .. Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswitch, MA, USA) following the manufacturer’s recommendations, and index codes were added to attribute the sequencing reads to each sample.

    DNA Methylation Assay:

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations. .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Concentration Assay:

    Article Title: Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells
    Article Snippet: RNA concentration was determined on Qubit® 2.0 Fluorometer (ThermoFisher/Invitrogen, Grand Island, NY), RNA quality was assessed using the Agilent 2100 Bioanalyzer. .. 2 μL of 1:2000 diluted External RNA Controls Consortium (ERCC) Spike-In Mix (Ambion™ 4,456,740, ThermoFisher Scientific Inc.) was added to 100 ng of high-quality total RNA followed by mRNA isolation using NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs E7490, Ipswich, MA) and RNA library construction with NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs E7530) according to the manufacturer’s user guide.

    Article Title: Mating-Induced Transcriptome Changes in the Reproductive Tract of Female Aedes aegypti
    Article Snippet: The concentration of RNA in each sample was measured using a Qubit spectrophotometer (Invitrogen, Grand Island, NY), and the quality of RNA was measured on a Fragment Analyzer (Advanced Analytical Technologies, Inc., Ames, IA). .. From total RNA, mRNA was extracted using the poly-A mRNA Magnetic Isolation Module (New England Biolabs, Inc., Ipswich, MA). cDNA libraries for each replicate were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Inc., Ipswich, MA).

    Article Title: In Utero Heat Stress Alters the Offspring Epigenome
    Article Snippet: Briefly, RNA was extracted from mammary tissue using the RNeasy Plus Universal Mini Kit (Qiagen, Valencia, CA) and RNA concentration was determined using a NanoDrop instrument (ND-2000, ThermoFisher Scientific, Waltham, MA). .. RNA-Seq library was constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) following manufacturer’s recommendations.

    Article Title: Studying on the strictly self-compatibility mechanism of ‘Liuyefeitao’ peach (Prunus persica L.)
    Article Snippet: The RNA concentration and purity were checked by OD A260/A280 ( > 1.8) and A260/A230 ( > 1.6), and the yield and quality were accessed using Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) and RNA 6000 Nano LabChip Kit (Agilent, CA, USA), RIN > 7.0. .. Transcriptome libraries were constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (New England Biolabs) according to the manufacturer’s instructions.

    Article Title: Functional Insights into the Roles of Hormones in the Dendrobium officinale-Tulasnella sp. Germinated Seed Symbiotic Association
    Article Snippet: RNA concentration was measured with the Qubit® RNA Assay Kit in the Qubit® 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA). .. Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswitch, MA, USA) following the manufacturer’s recommendations, and index codes were added to attribute the sequencing reads to each sample.

    Article Title: Comparative transcriptomics identifies genes differentially expressed in the intestine of a new fast-growing strain of common carp with higher unsaturated fatty acid content in muscle
    Article Snippet: To ensure that foregut, midgut and hindgut segments are represented in the transcriptome, RNA was extracted (Trizol, Invitrogen, US) separately from the three segments, and equal amounts (of the three segments belonging to the same specimen) were then pooled together. .. RNA integrity and concentration were assessed using an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). cDNA libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer’s recommendations. .. One μg of RNA per sample was used as input material.

    Lysis:

    Article Title: Small non-coding RNA landscape of extracellular vesicles from human stem cells
    Article Snippet: Before precipitation, two centrifugation steps were used to remove cellular components and two supernantant removal steps to get rid of the un-precipitated material, followed by EV lysis and RNA isolation using miRCURY™ RNA Isolation Kits - Cell & Plant (Exiqon A/S). .. NGS libraries were prepared using the NEBNext® Small RNA Library preparation kit (New England Biolabs), consisting of adapter ligation, cDNA conversion, PCR amplification (18 cycles) and purification.

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  • 99
    New England Biolabs multiplex small rna library prep kit
    Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from <t>immunoprecipitated</t> MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small <t>RNA</t> libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries
    Multiplex Small Rna Library Prep Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/multiplex small rna library prep kit/product/New England Biolabs
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    multiplex small rna library prep kit - by Bioz Stars, 2019-12
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    84
    New England Biolabs next srna kit
    Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from <t>immunoprecipitated</t> MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small <t>RNA</t> libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries
    Next Srna Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/next srna kit/product/New England Biolabs
    Average 84 stars, based on 1 article reviews
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    Image Search Results


    Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small RNA libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries

    Journal: Nature Communications

    Article Title: PNLDC1 is essential for piRNA 3′ end trimming and transposon silencing during spermatogenesis in mice

    doi: 10.1038/s41467-017-00854-4

    Figure Lengend Snippet: Increased piRNA sizes and reduced normal piRNAs in adult Pnldc1 Mut testes. a piRNA extension in Pnldc1 Mut mice. Total RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testes were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Square bracket indicates extended piRNAs. b MILI-piRNA extension in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MILI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. c MIWI-piRNA extension and reduction in Pnldc1 Mut (Mut-1) mice. Small RNAs were isolated from immunoprecipitated MIWI RNPs and were end-labeled with [ 32 P]-ATP, and detected by 15% TBE urea gel and autoradiography. Western blotting was performed with anti-MIWI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M molecular weight marker. d The length distribution of small RNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) testicular small RNA libraries. Data were normalized by microRNA reads (21–23 nt). e The length distribution of MILI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MILI-piRNA libraries. f The length distribution of MIWI-piRNAs from adult WT and Pnldc1 Mut (Mut-1 and Mut-2) MIWI-piRNA libraries

    Article Snippet: Small RNA libraries from immunoprecipitated RNAs or total RNA were prepared using NEBNext Multiplex Small RNA Library Prep Kit (E7300, NEB) following manufacturer’s instructions.

    Techniques: Mouse Assay, Labeling, Autoradiography, Isolation, Immunoprecipitation, Western Blot, Molecular Weight, Marker

    Increased piRNA lengths and reduced piRNA levels in adult Tdrkh cKO testes. ( A ) Severe reduction of total piRNAs in Tdrkh cKO testes. Total RNA from adult WT and Tdrkh cKO testes were end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. ( B ) The length distribution of small RNAs from adult WT and Tdrkh cKO testicular small RNA libraries. Data were normalized by miRNA reads (21–23nt). ( C ) Extension of MILI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MILI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M: molecular weight marker. ( D ) Diminished MIWI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MIWI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MIWI antibody to show immunoprecipitation efficiency. M: molecular weight marker. ( E ) The length distribution of MILI-piRNAs from adult WT and Tdrkh cKO MILI-piRNA libraries. ( F ) The length distribution of MIWI-piRNAs from adult WT and Tdrkh cKO MIWI-piRNA libraries.

    Journal: Nucleic Acids Research

    Article Title: Mitochondrial membrane-based initial separation of MIWI and MILI functions during pachytene piRNA biogenesis

    doi: 10.1093/nar/gky1281

    Figure Lengend Snippet: Increased piRNA lengths and reduced piRNA levels in adult Tdrkh cKO testes. ( A ) Severe reduction of total piRNAs in Tdrkh cKO testes. Total RNA from adult WT and Tdrkh cKO testes were end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. ( B ) The length distribution of small RNAs from adult WT and Tdrkh cKO testicular small RNA libraries. Data were normalized by miRNA reads (21–23nt). ( C ) Extension of MILI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MILI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MILI antibody to show immunoprecipitation efficiency. Square bracket indicates extended piRNAs. M: molecular weight marker. ( D ) Diminished MIWI-piRNAs in Tdrkh cKO testes. Small RNAs were isolated from immunoprecipitated MIWI-RNPs, end-labeled with [ 32 P]-ATP and detected by 15% TBE urea gel and autoradiography. Western blotting was performed using anti-MIWI antibody to show immunoprecipitation efficiency. M: molecular weight marker. ( E ) The length distribution of MILI-piRNAs from adult WT and Tdrkh cKO MILI-piRNA libraries. ( F ) The length distribution of MIWI-piRNAs from adult WT and Tdrkh cKO MIWI-piRNA libraries.

    Article Snippet: Small RNA libraries from immunoprecipitated RNAs or total RNA were prepared using NEBNext® Multiplex Small RNA Library Prep Kit (E7300, NEB) following manufacturer's instructions.

    Techniques: Labeling, Autoradiography, Isolation, Immunoprecipitation, Western Blot, Molecular Weight, Marker