mrna second strand synthesis module  (New England Biolabs)


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  • 99
    Name:
    NEBNext RNA First Strand Synthesis Module
    Description:
    NEBNext RNA First Strand Synthesis Module 96 rxns
    Catalog Number:
    E7525L
    Price:
    487
    Size:
    96 rxns
    Category:
    mRNA Template Preparation for PCR
    Score:
    85
    Buy from Supplier


    Structured Review

    New England Biolabs mrna second strand synthesis module
    NEBNext RNA First Strand Synthesis Module
    NEBNext RNA First Strand Synthesis Module 96 rxns
    https://www.bioz.com/result/mrna second strand synthesis module/product/New England Biolabs
    Average 99 stars, based on 42 article reviews
    Price from $9.99 to $1999.99
    mrna second strand synthesis module - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs). .. First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP. .. For optimizing the size selection, another round of size selection was performed, as described by Zhong et al. , in which the beads were used in 1.4 volumes of the cDNA solution.

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. The First strand and the second-strand cDNA were subsequently synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-) and DNA polymerase I and RNase H, respectively.

    Synthesized:

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X). .. Then, first strand cDNA was synthesized using M-MuLV Reverse Transcriptase(RNase H-)and random hexamer primers.

    Article Title: Genome-Wide Analysis of CDPK Family in Foxtail Millet and Determination of SiCDPK24 Functions in Drought Stress
    Article Snippet: The NanoPhotometer spectrophotometer (IMPLEN, Westlake Village, CA, United States) and the RNA 6000 Nano Assay Kit for the Agilent 2100 Bioanalyzer system (Agilent Technologies, Santa Clara, CA, United States) were used to assess RNA purity and integrity. .. After assessment, RNA purity was cleaved by using divalent cations in the NEBNext First Strand Synthesis reaction buffer under elevated temperature, and then first-strand cDNA was synthesized from the cleaved RNA fragments using reverse transcriptase and random primers. .. Subsequently, the double-stranded cDNA synthesis was carried out exploiting DNA polymerase I and RNase H. The cDNA library was prepared and then sequenced by Allwegene Technology Company in Beijing, China.

    Construct:

    Article Title: Comparative expression analysis identifies the respiratory transition-related miRNAs and their target genes in tissues of metamorphosing Chinese giant salamander (Andrias davidianus)
    Article Snippet: Sequencing libraries were constructed using 3 μg of RNA per sample using NEBNext®Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA), following the manufacturer’s instructions. .. Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Electrophoresis:

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: RNA degradation and contamination was monitored on 1% agarose gels electrophoresis. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Random Hexamer Labeling:

    Article Title: NTRK3 kinase fusions in Spitz tumours
    Article Snippet: Sequence reads were mapped to the reference human genome (hg19) using Tophat and fusions were identified by Tophat Fusion in combination with manual review [ , ]. .. Total RNA extracted from FFPE tissue sections was reverse transcribed with random hexamer primers using an NEBNext RNA First Strand Synthesis module. .. RT-PCR amplification was done for 32 cycles using Platinum Taq DNA polymerase (Invitrogen) with positive and negative controls using primer pairs described in .

    Formalin-fixed Paraffin-Embedded:

    Article Title: NTRK3 kinase fusions in Spitz tumours
    Article Snippet: Sequence reads were mapped to the reference human genome (hg19) using Tophat and fusions were identified by Tophat Fusion in combination with manual review [ , ]. .. Total RNA extracted from FFPE tissue sections was reverse transcribed with random hexamer primers using an NEBNext RNA First Strand Synthesis module. .. RT-PCR amplification was done for 32 cycles using Platinum Taq DNA polymerase (Invitrogen) with positive and negative controls using primer pairs described in .

    Activity Assay:

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature. .. First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Hybridization:

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X). .. Subsequently, second strand cDNA was synthesized using DNA Polymerase I and RNase H. The remaining overhangs were converted into blunt ends via exonuclease/polymerase activities.

    Article Title: Comparative expression analysis identifies the respiratory transition-related miRNAs and their target genes in tissues of metamorphosing Chinese giant salamander (Andrias davidianus)
    Article Snippet: Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Transcriptomic insights into the early host-pathogen interaction of cat intestine with Toxoplasma gondii
    Article Snippet: Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. First strand cDNA was synthesized using random hexamer primer and M-MuL V Reverse Transcriptase (RNase H− ).

    Article Title: Integrative Analyses of Transcriptome Sequencing Identify Functional miRNAs in the Chicken Embryo Fibroblasts Cells Infected With Reticuloendotheliosis Virus
    Article Snippet: Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×). .. Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×).

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Sequencing:

    Article Title: Diverse Long RNAs Are Differentially Sorted into Extracellular Vesicles Secreted by Colorectal Cancer Cells
    Article Snippet: Paragraph title: mRNA library preparation and sequencing ... First strand synthesis was performed using NEBNext RNA first strand synthesis module (New England BioLabs Inc.).

    Article Title: The life history of retrocopies illuminates the evolution of new mammalian genes
    Article Snippet: We additionally generated paired-end strand-specific RNA-seq data from orangutan testis (two sequencing experiments performed from the same cDNA library). .. The library was prepared using NEBNext RNA first-strand synthesis module (using Actinomycin D) and NEBNext ultra directional RNA second-strand synthesis module, and we performed two sequencing runs on the Illumina MiSeq platform. .. Reads were preprocessed by removing the last nucleotide at their 3′-end (artificially included by the protocol), and adaptors were trimmed using Skewer ( ).

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: In the present study, 3 μg of RNA per sample and the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) were used to generate the sequence libraries according to the manufacturer’s recommendations. .. In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X).

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Paragraph title: Preparation of strand-specific cDNA library for transcriptome sequencing ... Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP.

    Article Title: Comparative expression analysis identifies the respiratory transition-related miRNAs and their target genes in tissues of metamorphosing Chinese giant salamander (Andrias davidianus)
    Article Snippet: Paragraph title: Library preparation and transcriptome sequencing ... Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Genome-Wide Analysis of CDPK Family in Foxtail Millet and Determination of SiCDPK24 Functions in Drought Stress
    Article Snippet: After assessment, RNA purity was cleaved by using divalent cations in the NEBNext First Strand Synthesis reaction buffer under elevated temperature, and then first-strand cDNA was synthesized from the cleaved RNA fragments using reverse transcriptase and random primers. .. Subsequently, the double-stranded cDNA synthesis was carried out exploiting DNA polymerase I and RNase H. The cDNA library was prepared and then sequenced by Allwegene Technology Company in Beijing, China.

    Article Title: Transcriptomic insights into the early host-pathogen interaction of cat intestine with Toxoplasma gondii
    Article Snippet: Paragraph title: Library preparation for sequencing ... Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: Paragraph title: cDNA library preparation, clustering, and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Paragraph title: Sample preparation and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Paragraph title: cDNA library construction and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Paragraph title: cDNA library preparation and Illumina sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Sequencing libraries were generated using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Paragraph title: cDNA library construction and Illumina sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: Paragraph title: RNA Isolation and Sequencing Library Preparation ... First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Article Title: Integrative Analyses of Transcriptome Sequencing Identify Functional miRNAs in the Chicken Embryo Fibroblasts Cells Infected With Reticuloendotheliosis Virus
    Article Snippet: Paragraph title: Library construction and sequencing ... Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Under the direction of manufacturer, the sequencing libraries was prepared by using NEBNext® UltraTM RNA Library Prep Kit and index codes were added to attribute sequences to each sample. .. Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: Paragraph title: RNA preparation and library construction for transcriptome sequencing ... Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Ligation:

    Article Title: Diverse Long RNAs Are Differentially Sorted into Extracellular Vesicles Secreted by Colorectal Cancer Cells
    Article Snippet: First strand synthesis was performed using NEBNext RNA first strand synthesis module (New England BioLabs Inc.). .. First strand synthesis was performed using NEBNext RNA first strand synthesis module (New England BioLabs Inc.).

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs). .. First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP. .. Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP.

    Infection:

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Total RNAs were extracted individually from MDCC-MSB1 cells after infected with shRNA3-1657 or NC. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Generated:

    Article Title: The life history of retrocopies illuminates the evolution of new mammalian genes
    Article Snippet: We additionally generated paired-end strand-specific RNA-seq data from orangutan testis (two sequencing experiments performed from the same cDNA library). .. The library was prepared using NEBNext RNA first-strand synthesis module (using Actinomycin D) and NEBNext ultra directional RNA second-strand synthesis module, and we performed two sequencing runs on the Illumina MiSeq platform.

    Article Title: Transcriptomic insights into the early host-pathogen interaction of cat intestine with Toxoplasma gondii
    Article Snippet: Sequencing libraries were generated using an NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, Ipswich, USA) following manufacturer’s recommendations and index codes were added to correlate sequences to their respective samples. .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: The 1.5 μg total RNA per sample was used as input material for the RNA sample preparations, and sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Sequencing libraries were generated using a NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA) according to the manufacturer’s recommendations. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations, and mRNA purified from total RNA using poly-T oligo-linked magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Sequencing libraries were generated using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: A total amount of 1.5 µg RNA was used for preparing sequencing library generated by NEBNext® Ultra™ RNA Library Prep Kit. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: Sequencing libraries were generated using the NEBNext® UltraTM Directional RNA Library Prep Kit for Illumina® (NEB, Ipswich, MA, United States) according to the protocol. .. First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Article Title: Integrative Analyses of Transcriptome Sequencing Identify Functional miRNAs in the Chicken Embryo Fibroblasts Cells Infected With Reticuloendotheliosis Virus
    Article Snippet: After cluster generation, the library preparations were sequenced on an Illumina Hiseq 2500/2000 platform (Illumina) and 50 bp single-end reads were generated. .. Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Sequencing adaptors were ligated to cDNA fragments of preferentially 250~300 bp in length from each cDNA library, the ligated the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA) and enriched by PCR amplification.

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: Sequencing libraries were generated using a NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, Beverly, MA, USA), and index codes were added to attribute sequences to each sample. .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Polymerase Chain Reaction:

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs). .. First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: NTRK3 kinase fusions in Spitz tumours
    Article Snippet: Total RNA extracted from FFPE tissue sections was reverse transcribed with random hexamer primers using an NEBNext RNA First Strand Synthesis module. .. RT-PCR amplification was done for 32 cycles using Platinum Taq DNA polymerase (Invitrogen) with positive and negative controls using primer pairs described in .

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP. .. For optimizing the size selection, another round of size selection was performed, as described by Zhong et al. , in which the beads were used in 1.4 volumes of the cDNA solution.

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. After adenylation of 3′ ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization.

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. The First strand and the second-strand cDNA were subsequently synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-) and DNA polymerase I and RNase H, respectively.

    RNA Sequencing Assay:

    Article Title: Diverse Long RNAs Are Differentially Sorted into Extracellular Vesicles Secreted by Colorectal Cancer Cells
    Article Snippet: Approximately 500 ng of total RNA were required for proceeding to downstream RNA-seq applications. .. First strand synthesis was performed using NEBNext RNA first strand synthesis module (New England BioLabs Inc.).

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: Paragraph title: RNA-seq. ... First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: The life history of retrocopies illuminates the evolution of new mammalian genes
    Article Snippet: Paragraph title: RNA-seq data ... The library was prepared using NEBNext RNA first-strand synthesis module (using Actinomycin D) and NEBNext ultra directional RNA second-strand synthesis module, and we performed two sequencing runs on the Illumina MiSeq platform.

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: Paragraph title: Library preparation and RNA sequencing ... In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X).

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: For each sample, 5 μg of total RNA was used to isolate mRNA for the preparation of a strand-specific RNA-seq library, using a NEBNext Poly (A) mRNA Magnetic Isolation Module and NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) and following the manufacturer's protocols, with minor modifications. .. Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP.

    Article Title: Genome-Wide Analysis of CDPK Family in Foxtail Millet and Determination of SiCDPK24 Functions in Drought Stress
    Article Snippet: Paragraph title: RNA-Seq Analysis ... After assessment, RNA purity was cleaved by using divalent cations in the NEBNext First Strand Synthesis reaction buffer under elevated temperature, and then first-strand cDNA was synthesized from the cleaved RNA fragments using reverse transcriptase and random primers.

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Paragraph title: Library preparation for RNA-seq ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Paragraph title: RNA-Seq after linc-GALMD3 knockdown ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Paragraph title: 4.3. RNA Extraction and RNA-Seq ... Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Magnetic Beads:

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: In the present study, 3 μg of RNA per sample and the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) were used to generate the sequence libraries according to the manufacturer’s recommendations. .. In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X). .. Then, first strand cDNA was synthesized using M-MuLV Reverse Transcriptase(RNase H-)and random hexamer primers.

    Article Title: Comparative expression analysis identifies the respiratory transition-related miRNAs and their target genes in tissues of metamorphosing Chinese giant salamander (Andrias davidianus)
    Article Snippet: Index codes were added to attribute sequences to each sample. .. Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. First strand cDNA was transcribed using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H).

    Article Title: Transcriptomic insights into the early host-pathogen interaction of cat intestine with Toxoplasma gondii
    Article Snippet: The mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations, and mRNA purified from total RNA using poly-T oligo-linked magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Integrative Analyses of Transcriptome Sequencing Identify Functional miRNAs in the Chicken Embryo Fibroblasts Cells Infected With Reticuloendotheliosis Virus
    Article Snippet: Then, mRNAs were purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: We extracted the mRNA of each sample from the total RNA by using poly-T oligo-attached magnetic beads. .. Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Isolation:

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: The concentration and integrity of the isolated large (≥200-nt) RNA were estimated with a Qubit 2.0 Fluorometer (Invitrogen, Life Technologies, Inc.) and an Agilent 2100 Bioanalyzer (Applied Biosystems, Life Technologies, Inc.), respectively. .. First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: For each sample, 5 μg of total RNA was used to isolate mRNA for the preparation of a strand-specific RNA-seq library, using a NEBNext Poly (A) mRNA Magnetic Isolation Module and NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) and following the manufacturer's protocols, with minor modifications. .. Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP.

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: Paragraph title: RNA Isolation and Sequencing Library Preparation ... First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Purification:

    Article Title: Transcriptional profiling of two different physiological states of the yak mammary gland using RNA sequencing
    Article Snippet: In the present study, 3 μg of RNA per sample and the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) were used to generate the sequence libraries according to the manufacturer’s recommendations. .. In brief, after purification from total RNA using poly-T oligo-attached magnetic beads, mRNA was split into fragments using divalent cations under an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer(5X). .. Then, first strand cDNA was synthesized using M-MuLV Reverse Transcriptase(RNase H-)and random hexamer primers.

    Article Title: Comparative expression analysis identifies the respiratory transition-related miRNAs and their target genes in tissues of metamorphosing Chinese giant salamander (Andrias davidianus)
    Article Snippet: Index codes were added to attribute sequences to each sample. .. Briefly: mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. First strand cDNA was transcribed using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H).

    Article Title: Transcriptomic insights into the early host-pathogen interaction of cat intestine with Toxoplasma gondii
    Article Snippet: The mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations, and mRNA purified from total RNA using poly-T oligo-linked magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Long intergenic non-coding RNA GALMD3 in chicken Marek’s disease
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: A NanoPhotometer spectrophotometer (Implen; Westlake Village, CA, United States) was used to determine the quantity of purified RNA by measuring the absorbance at 260/280 nm. .. First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Article Title: Integrative Analyses of Transcriptome Sequencing Identify Functional miRNAs in the Chicken Embryo Fibroblasts Cells Infected With Reticuloendotheliosis Virus
    Article Snippet: Then, mRNAs were purified from total RNA using poly-T oligo-attached magnetic beads. .. Fragmentations were carried out using divalent cations under elevated temperature in NEBNext First-Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. The First strand and the second-strand cDNA were subsequently synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-) and DNA polymerase I and RNase H, respectively.

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: NTRK3 kinase fusions in Spitz tumours
    Article Snippet: Paragraph title: Confirmation by RT-PCR ... Total RNA extracted from FFPE tissue sections was reverse transcribed with random hexamer primers using an NEBNext RNA First Strand Synthesis module.

    cDNA Library Assay:

    Article Title: The life history of retrocopies illuminates the evolution of new mammalian genes
    Article Snippet: We additionally generated paired-end strand-specific RNA-seq data from orangutan testis (two sequencing experiments performed from the same cDNA library). .. The library was prepared using NEBNext RNA first-strand synthesis module (using Actinomycin D) and NEBNext ultra directional RNA second-strand synthesis module, and we performed two sequencing runs on the Illumina MiSeq platform.

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Paragraph title: Preparation of strand-specific cDNA library for transcriptome sequencing ... Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP.

    Article Title: Identification of Chemosensory Genes Based on the Transcriptomic Analysis of Six Different Chemosensory Organs in Spodoptera exigua
    Article Snippet: Paragraph title: cDNA library preparation, clustering, and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
    Article Snippet: Paragraph title: cDNA library construction and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext first strand synthesis reaction buffer (5X).

    Article Title: Sequencing and Characterization of the Invasive Sycamore Lace Bug Corythucha ciliata (Hemiptera: Tingidae) Transcriptome
    Article Snippet: Paragraph title: cDNA library preparation and Illumina sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: Paragraph title: cDNA library construction and Illumina sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×). .. The First strand and the second-strand cDNA were subsequently synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-) and DNA polymerase I and RNase H, respectively.

    RNA Extraction:

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Paragraph title: 4.3. RNA Extraction and RNA-Seq ... Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Selection:

    Article Title: De novo Assembly of the Camellia nitidissima Transcriptome Reveals Key Genes of Flower Pigment Biosynthesis
    Article Snippet: Briefly, mRNA was extracted with 15 μl of NEBNext Magnetic Oligo d(T)25 and fragmented in NEBNext First Strand Synthesis Buffer by heating at 94°C for 10 min. First-strand cDNA was reverse-transcribed from the fragmented mRNA and then used as a template to synthesize double-stranded cDNA, with dUTP replacing dTTP. .. To remove unwanted large fragments, the adaptor-ligated cDNA was selected for size using Agencourt AMPure XP beads (Beckman Coulter, Pasadena, CA, USA) in 0.6 volumes of the ligation reaction.

    Sample Prep:

    Article Title: hTERT promotes cell adhesion and migration independent of telomerase activity
    Article Snippet: Paragraph title: Sample preparation and sequencing ... Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X).

    Spectrophotometry:

    Article Title: Genome-Wide Analysis of CDPK Family in Foxtail Millet and Determination of SiCDPK24 Functions in Drought Stress
    Article Snippet: The NanoPhotometer spectrophotometer (IMPLEN, Westlake Village, CA, United States) and the RNA 6000 Nano Assay Kit for the Agilent 2100 Bioanalyzer system (Agilent Technologies, Santa Clara, CA, United States) were used to assess RNA purity and integrity. .. After assessment, RNA purity was cleaved by using divalent cations in the NEBNext First Strand Synthesis reaction buffer under elevated temperature, and then first-strand cDNA was synthesized from the cleaved RNA fragments using reverse transcriptase and random primers.

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: RNA purity was checked by using the NanoPhotometer® spectrophotometer (IMPLEN, CA, USA) and its concentration was measured using Qubit® RNA Assay Kit in Qubit® 2.0 Flurometer (Life Technologies, CA, USA). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: The NanoPhotometer® spectrophotometer was used to check RNA purity and concentration. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury
    Article Snippet: A NanoPhotometer spectrophotometer (Implen; Westlake Village, CA, United States) was used to determine the quantity of purified RNA by measuring the absorbance at 260/280 nm. .. First, performed by the divalent cations in NEBNext First Strand Synthesis Reaction Buffer at high temperature.

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: By using a NanoPhotometer® spectrophotometer (IMPLEN, Los Angeles, CA, USA), we checked the total RNA purity of each sample. .. Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: RNA purity was checked using the NanoPhotometer spectrophotometer (Implen, Inc., Westlake Village, CA, USA). .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Concentration Assay:

    Article Title: Diverse Long RNAs Are Differentially Sorted into Extracellular Vesicles Secreted by Colorectal Cancer Cells
    Article Snippet: The concentration and integrity of the extracted total RNA was estimated by Qubit® 2.0 Fluorometer (Invitrogen), and Agilent 2100 Bioanalyzer (Applied Biosystems), respectively. .. First strand synthesis was performed using NEBNext RNA first strand synthesis module (New England BioLabs Inc.).

    Article Title: Modulation of microRNA-mRNA Target Pairs by Human Papillomavirus 16 Oncoproteins
    Article Snippet: The concentration and integrity of the isolated large (≥200-nt) RNA were estimated with a Qubit 2.0 Fluorometer (Invitrogen, Life Technologies, Inc.) and an Agilent 2100 Bioanalyzer (Applied Biosystems, Life Technologies, Inc.), respectively. .. First, rRNA was removed with the Ribo-Zero Magnetic Gold (Yeast) kit (Epicentre, Illumina) according to the manufacturer’s recommended protoco l. The RNA was then fragmented and primed for first-strand synthesis with the NEBNext RNA First Strand Synthesis Module (New England Biolabs).

    Article Title: RNA-seq comparative analysis of Peking ducks spleen gene expression 24 h post-infected with duck plague virulent or attenuated virus
    Article Snippet: RNA purity was checked by using the NanoPhotometer® spectrophotometer (IMPLEN, CA, USA) and its concentration was measured using Qubit® RNA Assay Kit in Qubit® 2.0 Flurometer (Life Technologies, CA, USA). .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Identification of the pheromone biosynthesis genes from the sex pheromone gland transcriptome of the diamondback moth, Plutella xylostella
    Article Snippet: The NanoPhotometer® spectrophotometer was used to check RNA purity and concentration. .. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Comparative Analysis of the Integument Transcriptomes between stick Mutant and Wild-Type Silkworms
    Article Snippet: Then, the Qubit® RNA Assay Kit in Qubit® 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, Palo Alto, CA, USA) were used to determine the RNA concentration and integrity, respectively. .. Then, the mRNA was fragmented by the divalent cation with an elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Article Title: Transcriptome profiling reveals the role of ZBTB38 knock-down in human neuroblastoma
    Article Snippet: RNA concentration was measured using the Qubit RNA Assay Kit in Qubit 2.0 Fluorometer (Life Technologies, Camarillo, CA, USA). .. Fragmentation was performed using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5×).

    Staining:

    Article Title: NTRK3 kinase fusions in Spitz tumours
    Article Snippet: Total RNA extracted from FFPE tissue sections was reverse transcribed with random hexamer primers using an NEBNext RNA First Strand Synthesis module. .. RT-PCR amplification was done for 32 cycles using Platinum Taq DNA polymerase (Invitrogen) with positive and negative controls using primer pairs described in .

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