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    Illumina Inc illumina hiseq 2500
    Experimental scheme. ( A ) After differentiation, the cell cultures were dissociated to single-cell suspensions and loaded onto a Fluidigm C1 Single-Cell Auto Prep Array for mRNA-Seq. On the arrays, the cells were lysed, and reverse transcription of the mRNA and PCR amplification of the cDNA were performed using the C1 Single-Cell Auto Prep System (Fluidigm). Libraries were then prepared using the Nextera XT DNA Library Prep Kit and mosquito HTS liquid handler (TTP). For the final PCR reactions, we used a Bio-Rad 384 Thermal Cycler. Libraries were pooled and sequenced on an <t>Illumina</t> HiSeq 2500. ( B ) WA09 human embryonic stem cells were differentiated in vitro to the pancreatic lineage. Cells from stages 1 and 2 were collected and analyzed using the procedures outlined in A . Two independent cells from stage 1 (cell A and cell B) and two cells from stage 2 (cell C and cell D) were selected and yielded with similar cDNA concentrations (mean [SD] concentration = 0.38 [0.04] ng/µL). For library preparation, we tested 2-µL, 4-µL, and 8-µL final volume reactions, with four replicates per reaction volume.
    Illumina Hiseq 2500, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 99/100, based on 31876 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Illumina Inc hiseq 2500 sequencer
    Performance comparisons of BBT against FACS, BWA and BT2. Receiver operator characteristic curves of BBT and FACS using simulated 100 bp SE reads from Homo sapiens mixed with ( A ) E.coli and ( B ) Mus musculus filtered against an H.sapiens Bloom filter using a k -mer size of 25 bp; ( C ) CPU time benchmark comparing BT2 (for a range of built-in settings), BWA (using aln and mem settings), FACS and BBT, on one lane of human 2 × 150 bp PE Illumina <t>HiSeq</t> 2500 reads
    Hiseq 2500 Sequencer, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 93/100, based on 4757 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiseq 2500 sequencer/product/Illumina Inc
    Average 93 stars, based on 4757 article reviews
    Price from $9.99 to $1999.99
    hiseq 2500 sequencer - by Bioz Stars, 2020-09
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    92
    Illumina Inc hiseq 2500 sequencing system
    Illustrated representation of the optimized barcoded cDNA library preparation procedure. Total RNA from formalin-fixed paraffin-embedded (FFPE) specimens ( n = 18), are ligated with 3′ adapters ( n = 18), purified on a 15% PAGE gel (using size markers (SM) RNA oligonucleotides) and excised after SYBR ® gold staining of the gel. The combined purified products are ligated with a 5′ adapter, run on a 12% PAGE gel, excised and purified. The dual-adapter ligated small-RNA products are reverse transcribed, subjected to a pilot PCR for identification of adequate amplification cycle. A large-scale PCR is performed, and the products are separated on a 2% agarose gel, where the upper band (cDNA library) is purified and subjected to sequencing on an Illumina <t>HiSeq-2500</t> sequencer.
    Hiseq 2500 Sequencing System, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 1669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiseq 2500 sequencing system/product/Illumina Inc
    Average 92 stars, based on 1669 article reviews
    Price from $9.99 to $1999.99
    hiseq 2500 sequencing system - by Bioz Stars, 2020-09
    92/100 stars
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    90
    Illumina Inc hiseq 2000 2500 platform
    Illustrated representation of the optimized barcoded cDNA library preparation procedure. Total RNA from formalin-fixed paraffin-embedded (FFPE) specimens ( n = 18), are ligated with 3′ adapters ( n = 18), purified on a 15% PAGE gel (using size markers (SM) RNA oligonucleotides) and excised after SYBR ® gold staining of the gel. The combined purified products are ligated with a 5′ adapter, run on a 12% PAGE gel, excised and purified. The dual-adapter ligated small-RNA products are reverse transcribed, subjected to a pilot PCR for identification of adequate amplification cycle. A large-scale PCR is performed, and the products are separated on a 2% agarose gel, where the upper band (cDNA library) is purified and subjected to sequencing on an Illumina <t>HiSeq-2500</t> sequencer.
    Hiseq 2000 2500 Platform, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 90/100, based on 441 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiseq 2000 2500 platform/product/Illumina Inc
    Average 90 stars, based on 441 article reviews
    Price from $9.99 to $1999.99
    hiseq 2000 2500 platform - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    91
    Illumina Inc illumina hiseq 2000 2500
    Illustrated representation of the optimized barcoded cDNA library preparation procedure. Total RNA from formalin-fixed paraffin-embedded (FFPE) specimens ( n = 18), are ligated with 3′ adapters ( n = 18), purified on a 15% PAGE gel (using size markers (SM) RNA oligonucleotides) and excised after SYBR ® gold staining of the gel. The combined purified products are ligated with a 5′ adapter, run on a 12% PAGE gel, excised and purified. The dual-adapter ligated small-RNA products are reverse transcribed, subjected to a pilot PCR for identification of adequate amplification cycle. A large-scale PCR is performed, and the products are separated on a 2% agarose gel, where the upper band (cDNA library) is purified and subjected to sequencing on an Illumina <t>HiSeq-2500</t> sequencer.
    Illumina Hiseq 2000 2500, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 91/100, based on 409 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/illumina hiseq 2000 2500/product/Illumina Inc
    Average 91 stars, based on 409 article reviews
    Price from $9.99 to $1999.99
    illumina hiseq 2000 2500 - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    Experimental scheme. ( A ) After differentiation, the cell cultures were dissociated to single-cell suspensions and loaded onto a Fluidigm C1 Single-Cell Auto Prep Array for mRNA-Seq. On the arrays, the cells were lysed, and reverse transcription of the mRNA and PCR amplification of the cDNA were performed using the C1 Single-Cell Auto Prep System (Fluidigm). Libraries were then prepared using the Nextera XT DNA Library Prep Kit and mosquito HTS liquid handler (TTP). For the final PCR reactions, we used a Bio-Rad 384 Thermal Cycler. Libraries were pooled and sequenced on an Illumina HiSeq 2500. ( B ) WA09 human embryonic stem cells were differentiated in vitro to the pancreatic lineage. Cells from stages 1 and 2 were collected and analyzed using the procedures outlined in A . Two independent cells from stage 1 (cell A and cell B) and two cells from stage 2 (cell C and cell D) were selected and yielded with similar cDNA concentrations (mean [SD] concentration = 0.38 [0.04] ng/µL). For library preparation, we tested 2-µL, 4-µL, and 8-µL final volume reactions, with four replicates per reaction volume.

    Journal: Jala (Charlottesville, Va.)

    Article Title: Miniaturization Technologies for Efficient Single-Cell Library Preparation for Next-Generation Sequencing

    doi: 10.1177/2211068216630741

    Figure Lengend Snippet: Experimental scheme. ( A ) After differentiation, the cell cultures were dissociated to single-cell suspensions and loaded onto a Fluidigm C1 Single-Cell Auto Prep Array for mRNA-Seq. On the arrays, the cells were lysed, and reverse transcription of the mRNA and PCR amplification of the cDNA were performed using the C1 Single-Cell Auto Prep System (Fluidigm). Libraries were then prepared using the Nextera XT DNA Library Prep Kit and mosquito HTS liquid handler (TTP). For the final PCR reactions, we used a Bio-Rad 384 Thermal Cycler. Libraries were pooled and sequenced on an Illumina HiSeq 2500. ( B ) WA09 human embryonic stem cells were differentiated in vitro to the pancreatic lineage. Cells from stages 1 and 2 were collected and analyzed using the procedures outlined in A . Two independent cells from stage 1 (cell A and cell B) and two cells from stage 2 (cell C and cell D) were selected and yielded with similar cDNA concentrations (mean [SD] concentration = 0.38 [0.04] ng/µL). For library preparation, we tested 2-µL, 4-µL, and 8-µL final volume reactions, with four replicates per reaction volume.

    Article Snippet: Sequencing The 48 pooled libraries were sequenced on one lane of an Illumina HiSeq 2500 at an average total read depth of 5.6 million reads per sample (ST3).

    Techniques: Polymerase Chain Reaction, Amplification, In Vitro, Concentration Assay

    Schematic work-flow of C. retropictus transcriptome using Illumina sequencing, de novo analysis, and annotation. The visceral mass transcriptome of C. retropictus was obtained using an Illumina HiSeq 2500 platform. The raw reads were pre-processed using the Sickle software tool (quality: 20, length: 40) and Fastq_filter software to obtain clean reads. Using Trinity (K-mer: 25; minimum contig length: 200) de novo assembler and TGICL clustering, the clean reads were processed to unigene sequences. Subsequently, the unigene sequences were blasted against public databases including PANM, Unigene, COG, GO, and KEGG for functional annotation. The SSRs were detected within the unigene sequences using MISA software.

    Journal: Genes

    Article Title: Transcriptomic Analysis of the Endangered Neritid Species Clithon retropictus: De Novo Assembly, Functional Annotation, and Marker Discovery

    doi: 10.3390/genes7070035

    Figure Lengend Snippet: Schematic work-flow of C. retropictus transcriptome using Illumina sequencing, de novo analysis, and annotation. The visceral mass transcriptome of C. retropictus was obtained using an Illumina HiSeq 2500 platform. The raw reads were pre-processed using the Sickle software tool (quality: 20, length: 40) and Fastq_filter software to obtain clean reads. Using Trinity (K-mer: 25; minimum contig length: 200) de novo assembler and TGICL clustering, the clean reads were processed to unigene sequences. Subsequently, the unigene sequences were blasted against public databases including PANM, Unigene, COG, GO, and KEGG for functional annotation. The SSRs were detected within the unigene sequences using MISA software.

    Article Snippet: Finally, the library preparations were sequenced on an Illumina HiSeq 2500 platform at the sequencing facility of GnC Company (Daejeon, South Korea) with the generation of 100-base pair (bp) PE-reads.

    Techniques: Flow Cytometry, Sequencing, Software, Functional Assay

    (a) Hierarchical cluster analysis for genes included in non-fluorescent liposomes. Ten non-fluorescent liposomes that did not show β-galactoside hydrolysis activity were isolated, and the genes included in these liposomes were determined by multiplex next-generation sequencing using HiSeq 2500. In the hierarchical cluster analysis for these genes, there was no commonality among the genes included in each liposome, which indicated a random pattern. Each row represents a separate gene, each column represents a separate liposome and genes found in each liposome are shown in red. (b) Hierarchical cluster analysis for the genes included in fluorescent liposomes. Hierarchical cluster analysis revealed that  LacZ  was a clear cluster that was included in all fluorescent liposomes and with no other common factors.

    Journal: Scientific Reports

    Article Title: Integrating reductive and synthetic approaches in biology using man-made cell-like compartments

    doi: 10.1038/srep04722

    Figure Lengend Snippet: (a) Hierarchical cluster analysis for genes included in non-fluorescent liposomes. Ten non-fluorescent liposomes that did not show β-galactoside hydrolysis activity were isolated, and the genes included in these liposomes were determined by multiplex next-generation sequencing using HiSeq 2500. In the hierarchical cluster analysis for these genes, there was no commonality among the genes included in each liposome, which indicated a random pattern. Each row represents a separate gene, each column represents a separate liposome and genes found in each liposome are shown in red. (b) Hierarchical cluster analysis for the genes included in fluorescent liposomes. Hierarchical cluster analysis revealed that LacZ was a clear cluster that was included in all fluorescent liposomes and with no other common factors.

    Article Snippet: Next, the liposome-derived DNA fragments and the E. coli ORF library were pre-treated for HiSeq 2500 according to the Nextera XT DNA preparation kit protocol (Illumina).

    Techniques: Activity Assay, Isolation, Multiplex Assay, Next-Generation Sequencing

    MiRNA expressions profiles of exosomes. (A) Profiling of small RNAs in exosome samples. (B) A Venn diagram showing the co-expressed and specifically expressed miRNAs in exosomes. (C) Heat map of sequencing results. Gene expression data obtained using next-generation sequencing on the Illumina HiSeq 2500 platform. P

    Journal: Translational Oncology

    Article Title: Circulating Exosomal miR-17-5p and miR-92a-3p Predict Pathologic Stage and Grade of Colorectal Cancer

    doi: 10.1016/j.tranon.2017.12.012

    Figure Lengend Snippet: MiRNA expressions profiles of exosomes. (A) Profiling of small RNAs in exosome samples. (B) A Venn diagram showing the co-expressed and specifically expressed miRNAs in exosomes. (C) Heat map of sequencing results. Gene expression data obtained using next-generation sequencing on the Illumina HiSeq 2500 platform. P

    Article Snippet: To investigate the molecular mechanism of primary and metastatic CRC, we compared the miRNA expression profiles of exosomes secreted by two isogenic human CRC cell lines (primary SW480 cell line and its lymph node metastatic variant SW620) using the Illumina HiSeq 2500 system.

    Techniques: Sequencing, Expressing, Next-Generation Sequencing

    Measuring technical variation using DNA ladders. a Density distribution of k-mer counts for each cn unit ( n = 570 k-mers per unit; 1 cn = red, 2 cn = yellow, 4 cn = green, and 8 cn = blue) in a simulated and experimental libraries (prepared with KAPA HyperPlus PCR-free and sequenced on Illumina HiSeq X Ten), with the mean, standard deviation and coefficient of variation indicated. For the experimental library, the n = 14 independent synthetic ladders where examined over 5 independent experiments with comparable results (see HiSeq X Ten/PCR-free libraries in Supplementary Figs. 5 , 6 , and 7a ). b Modeling the impact of common NGS technical variables such as error rate, library depth, and library complexity on the regression slope of the synthetic DNA ladder. c The panel illustrates the variability associated with cn units across a range of libraries generated with different sequencing technologies (e.g., ONT Nanopore MinION, ONT Nanopore PromethION, Illumina HiSeq X Ten, Illumina NextSeq 500, and Illumina HiSeq 2500 instruments) or prepared with alternative protocols (e.g., KAPA, KAPA HyperPlus PCR-free/PCR-based and Nextera XT kits, or target enrichment by oligonucleotide hybridization). The y axis indicates k-mer counts, whilst the x axis indicates the position across the sub-sequence length (600 nt). The mean k-mer count (opaque line) and standard deviation (transparent line) are determined across all n = 14 independent synthetic ladders in all the sequencing experiments.

    Journal: Nature Communications

    Article Title: A universal and independent synthetic DNA ladder for the quantitative measurement of genomic features

    doi: 10.1038/s41467-020-17445-5

    Figure Lengend Snippet: Measuring technical variation using DNA ladders. a Density distribution of k-mer counts for each cn unit ( n = 570 k-mers per unit; 1 cn = red, 2 cn = yellow, 4 cn = green, and 8 cn = blue) in a simulated and experimental libraries (prepared with KAPA HyperPlus PCR-free and sequenced on Illumina HiSeq X Ten), with the mean, standard deviation and coefficient of variation indicated. For the experimental library, the n = 14 independent synthetic ladders where examined over 5 independent experiments with comparable results (see HiSeq X Ten/PCR-free libraries in Supplementary Figs. 5 , 6 , and 7a ). b Modeling the impact of common NGS technical variables such as error rate, library depth, and library complexity on the regression slope of the synthetic DNA ladder. c The panel illustrates the variability associated with cn units across a range of libraries generated with different sequencing technologies (e.g., ONT Nanopore MinION, ONT Nanopore PromethION, Illumina HiSeq X Ten, Illumina NextSeq 500, and Illumina HiSeq 2500 instruments) or prepared with alternative protocols (e.g., KAPA, KAPA HyperPlus PCR-free/PCR-based and Nextera XT kits, or target enrichment by oligonucleotide hybridization). The y axis indicates k-mer counts, whilst the x axis indicates the position across the sub-sequence length (600 nt). The mean k-mer count (opaque line) and standard deviation (transparent line) are determined across all n = 14 independent synthetic ladders in all the sequencing experiments.

    Article Snippet: They were then sequenced on a HiSeq 2500 instrument (Illumina®) producing paired reads of 125 nt at the Kinghorn Centre for Clinical Genomics, Sydney, Australia.

    Techniques: Polymerase Chain Reaction, Standard Deviation, Next-Generation Sequencing, Generated, Sequencing, Hybridization

    Work flow used to identify somatic variants in genomic DNA extracted from human lens epithelial cells. Sequencing libraries were enriched for genes of the WUCaMP2 panel ( Table 2 ) by targeted hybridization capture. The resulting library was sequenced on an Illumina HiSeq 2500 platform to obtain paired-end reads. Sequencing results were filtered at several levels. The reads were then aligned to a human reference genome (hg19), and PCR duplicates were removed. Variants were called using SAMtools and VarScan2 software. Finally, selected variants were inspected manually using the IGV visualization tool.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Somatic Variants in the Human Lens Epithelium: A Preliminary Assessment

    doi: 10.1167/iovs.16-19726

    Figure Lengend Snippet: Work flow used to identify somatic variants in genomic DNA extracted from human lens epithelial cells. Sequencing libraries were enriched for genes of the WUCaMP2 panel ( Table 2 ) by targeted hybridization capture. The resulting library was sequenced on an Illumina HiSeq 2500 platform to obtain paired-end reads. Sequencing results were filtered at several levels. The reads were then aligned to a human reference genome (hg19), and PCR duplicates were removed. Variants were called using SAMtools and VarScan2 software. Finally, selected variants were inspected manually using the IGV visualization tool.

    Article Snippet: Sequencing and Base Calling Multiplex sequencing of the DNA libraries was performed on a HiSeq 2500 sequencing platform (Illumina Inc., San Diego, CA, USA) to obtain paired end 101-bp reads.

    Techniques: Flow Cytometry, Sequencing, Hybridization, Polymerase Chain Reaction, Software

    ( A ) Schematic representation of metagenomic sequencing: BAL was split into two aliquots which were independently treated with two DNA extraction protocols: Ultra-Deep Microbiome Prep (Molzym) and NucleoSpin Soil (MN). Metagenomic libraries were independently sequenced with 2 × 250 HiSeq 2500 and 2 × 100 Hiseq 2000, respectively; ( B ) Schematic representation of bioinformatics analyses: forward and reverse raw reads of Molzym- (HiSeq Molzym (2 × 250)) and MN-treated (HiSeq MN (2 × 100)) samples were used for taxonomic analysis with CLARK, Kraken and MetaPhlAn2. In addition, before taxonomic analyses, read pairs of the HiSeq Molzym (2 × 250) dataset were either quality-filtered and merged (HiSeq Molzym merged) or trimmed to the length of 100 nt (HiSeq Molzym trimmed to 2 × 100) as described in Materials and Methods; ( C ) Pie-charts representing the proportions (%) of sequencing reads classified as human (in blue), prokaryotic (red) or unclassified (grey) by CLARK (top row) and Kraken (bottom row). Proportions were computed over the total number of reads present in a given dataset. Viral- and fungal-classified read proportions are not shown and contributed to less 0.02% of total reads in all sequencing datasets.

    Journal: International Journal of Molecular Sciences

    Article Title: Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing

    doi: 10.3390/ijms18092011

    Figure Lengend Snippet: ( A ) Schematic representation of metagenomic sequencing: BAL was split into two aliquots which were independently treated with two DNA extraction protocols: Ultra-Deep Microbiome Prep (Molzym) and NucleoSpin Soil (MN). Metagenomic libraries were independently sequenced with 2 × 250 HiSeq 2500 and 2 × 100 Hiseq 2000, respectively; ( B ) Schematic representation of bioinformatics analyses: forward and reverse raw reads of Molzym- (HiSeq Molzym (2 × 250)) and MN-treated (HiSeq MN (2 × 100)) samples were used for taxonomic analysis with CLARK, Kraken and MetaPhlAn2. In addition, before taxonomic analyses, read pairs of the HiSeq Molzym (2 × 250) dataset were either quality-filtered and merged (HiSeq Molzym merged) or trimmed to the length of 100 nt (HiSeq Molzym trimmed to 2 × 100) as described in Materials and Methods; ( C ) Pie-charts representing the proportions (%) of sequencing reads classified as human (in blue), prokaryotic (red) or unclassified (grey) by CLARK (top row) and Kraken (bottom row). Proportions were computed over the total number of reads present in a given dataset. Viral- and fungal-classified read proportions are not shown and contributed to less 0.02% of total reads in all sequencing datasets.

    Article Snippet: Pre-Processing of Sequencing Data Paired-ended sequencing of DNA preparations obtained without (NucleoSpin Soil kit, Macherey-Nagel, Düren, Germany) and with bacterial/fungal DNA enrichment (Ultra-Deep Microbiome Prep kit, Molzym, Bremen, Germany) was performed by Illumina HiSeq 2000 and by Illumina HiSeq 2500 platforms ( A), respectively.

    Techniques: Sequencing, DNA Extraction

    Performance comparisons of BBT against FACS, BWA and BT2. Receiver operator characteristic curves of BBT and FACS using simulated 100 bp SE reads from Homo sapiens mixed with ( A ) E.coli and ( B ) Mus musculus filtered against an H.sapiens Bloom filter using a k -mer size of 25 bp; ( C ) CPU time benchmark comparing BT2 (for a range of built-in settings), BWA (using aln and mem settings), FACS and BBT, on one lane of human 2 × 150 bp PE Illumina HiSeq 2500 reads

    Journal: Bioinformatics

    Article Title: BioBloom tools: fast, accurate and memory-efficient host species sequence screening using bloom filters

    doi: 10.1093/bioinformatics/btu558

    Figure Lengend Snippet: Performance comparisons of BBT against FACS, BWA and BT2. Receiver operator characteristic curves of BBT and FACS using simulated 100 bp SE reads from Homo sapiens mixed with ( A ) E.coli and ( B ) Mus musculus filtered against an H.sapiens Bloom filter using a k -mer size of 25 bp; ( C ) CPU time benchmark comparing BT2 (for a range of built-in settings), BWA (using aln and mem settings), FACS and BBT, on one lane of human 2 × 150 bp PE Illumina HiSeq 2500 reads

    Article Snippet: 3.2 Benchmarking on experimental data We used a single lane of 2 × 150 bp PE human DNA reads ( https://basespace.illumina.com/run/716717/2x150-HiSeq-2500-demo-NA12878 ) generated with an Illumina HiSeq 2500 sequencer to benchmark computational performance.

    Techniques: FACS

    Illustrated representation of the optimized barcoded cDNA library preparation procedure. Total RNA from formalin-fixed paraffin-embedded (FFPE) specimens ( n = 18), are ligated with 3′ adapters ( n = 18), purified on a 15% PAGE gel (using size markers (SM) RNA oligonucleotides) and excised after SYBR ® gold staining of the gel. The combined purified products are ligated with a 5′ adapter, run on a 12% PAGE gel, excised and purified. The dual-adapter ligated small-RNA products are reverse transcribed, subjected to a pilot PCR for identification of adequate amplification cycle. A large-scale PCR is performed, and the products are separated on a 2% agarose gel, where the upper band (cDNA library) is purified and subjected to sequencing on an Illumina HiSeq-2500 sequencer.

    Journal: International Journal of Molecular Sciences

    Article Title: Evaluation and Adaptation of a Laboratory-Based cDNA Library Preparation Protocol for Retrospective Sequencing of Archived MicroRNAs from up to 35-Year-Old Clinical FFPE Specimens

    doi: 10.3390/ijms18030627

    Figure Lengend Snippet: Illustrated representation of the optimized barcoded cDNA library preparation procedure. Total RNA from formalin-fixed paraffin-embedded (FFPE) specimens ( n = 18), are ligated with 3′ adapters ( n = 18), purified on a 15% PAGE gel (using size markers (SM) RNA oligonucleotides) and excised after SYBR ® gold staining of the gel. The combined purified products are ligated with a 5′ adapter, run on a 12% PAGE gel, excised and purified. The dual-adapter ligated small-RNA products are reverse transcribed, subjected to a pilot PCR for identification of adequate amplification cycle. A large-scale PCR is performed, and the products are separated on a 2% agarose gel, where the upper band (cDNA library) is purified and subjected to sequencing on an Illumina HiSeq-2500 sequencer.

    Article Snippet: The cDNA libraries were then sequenced (single-read 50 cycles) on a HiSeq 2500 Sequencing System (SY-401-2501, Illumina, San Diego, CA, USA).

    Techniques: cDNA Library Assay, Formalin-fixed Paraffin-Embedded, Purification, Polyacrylamide Gel Electrophoresis, Staining, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Sequencing