Small RNA Library Prep Kit for Illumina All the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Small RNA Library Prep Kit for Illumina All the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform Price USD$1,635.00 SKU 63600 Format Indexes 1-24 Indexes 25-48 Size 24 Preps Quantity Overview Details Documentation Citations Overview Optimized for low input RNA, especially from bodily fluids such as plasma or serum at 1ng of RNASimple and quick workflow: library could be prepared in less than 5 hoursNo gel purification for selected types of samplesProtocol optimized for RNA isolated from different types of input, including liquid biopsies (blood, plasma, serum and urine)Complements Norgen's Best-in-Class Total RNA (including microRNA) Purification Technology The Small RNA Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform. All molecular reagents including adaptors, primers, enzyme mixes and buffers are provided. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patent resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.Workflow Details Supporting Data Previous Figure 1. An example of a purified small RNA library on an Agilent 2100 Bioanalyzer using a High Sensitivity DNA Chip. The library was prepared using a mixture of synthetic microRNAs as an input. A single peak of ~ 141 bp was obtained and could be used directly for analysis on an Illumina next-generation sequencing platform. Figure 2. An example of a small RNA library on resolved on a 6% Novex® TBE PAGE gel. The library containing microRNAs resolved at around 140 bp and migrated just below the 150 bp marker of the provided NGS MW Ladder. The NGS Control Ladder contains two bands for alignment during band excision. The top band co-migrate with the expected size of library construct containing mainly microRNAs (~140 bp). The bottom band co-migrate with the expected size of an adaptor-adaptor-ligation product (~ 120 bp) that should be excluded. Figure 3. Superb reproducibility of constructing small RNA (microRNA) sequencing libraries from liquid biopsies samples with low RNA abundance. Total RNA was prepared from 200 µL of EDTA-plasma using Norgen's Plasma/Serum RNA Purification Mini Kit (Cat. 55000). Small RNA libraries (four replicates) were prepared from purified RNA using Norgen’s Small RNA Library Prep Kit for Illumina (Cat. 63600) without the use of gel purification (prep time of ~ 5 hours). The resulting libraries were sequenced on Illumina's MiSeq system using v3 chemistry. Reads were mapped to microRNAs and normalized to reads per million. Pairwise-comparisons of microRNA expression among all replicates were performed. High correlations (or reproducibility) were observed among all replicates with an average of over 99%. Figure 4. Combining true total RNA purification technology with robust production of small RNA (microRNA) sequencing libraries from RNA purified from whole blood and preserved blood. Total RNA was prepared from either whole blood or collected in Paxgene™ or Tempus™ blood tubes using Norgen’s Total RNA Purification Plus Kit (Cat. 48300), Preserved Blood RNA Purification Kit II (for use with Paxgene™ Blood RNA Tubes) (Cat. 43500) or Preserved Blood RNA Purification Kit I (for use with Tempus™ Blood RNA Tubes) (Cat. 43400), respectively, in duplicates. Small RNA libraries were prepared from purified RNA using Norgen's Small RNA Library Prep Kit for Illumina (Cat. 63600). The resulting libraries were sequenced on Illumina's MiSeq system using v3 chemistry. Reads were mapped to microRNAs and normalized to reads per million. Pairwise-comparisons of microRNA expression among all replicates were performed. High correlations (or reproducibility) were observed among all replicates of each blood tube with an average of over 99% (Panel A). Moreover, all three types of blood samples yielded similar numbers of microRNA detected (Panel B) with highly correlated expression profiles (Panel C) Next Click for expanded view Kit Specifications Number of preps 24 Number of indices provided 12 (for 24 preps, indexes 1-24 or 25-48) Time required to complete 6 libraries As little as 5 hours Input RNA required As little as 50 pg Storage Conditions and Product Stability Some components require storage at -20°C, 4°C or room temperature. See individual components and box labels for storage conditions. Step Component Cat. 64600 (24 preps) 3' AdaptorLigation to Template RNA 3' Adaptor 30 µL 3' Adaptor Ligation Master Mix 320 µL T4 RNA Ligase 2 (Truncated) 35 µL 5' Adaptor Ligation 5' Adaptor 30 µL 5' Adaptor Ligation Master Mix 320 µL T4 RNA Ligase 1 35 µL cDNA Synthesis from Ligated RNA Product Reverse Primer 30 µL cDNA Synthesis Master Mix 220 µL TruScript ReverseTranscriptase 35 µL PCR Amplification 2x NGS PCR Master Mix 1.32 µL PCR Reverse Primer 81 µL Forward Index Primer Included in Small RNA Library Prep Forward Index Primers (# 64640 or # 64610) Size Selection NGS MW Ladder 50 µL NGS Control Ladder 50 µL Loading Dye 300 µL Nuclease-Free Water 1.25 µL Component Cat. 63500 (75 preps) Buffer RL 40 mL Wash Solution A 38 mL (Reconstituted to 128 mL) Elution Solution A 6 mL Columns 75 Gel Filtration Columns 24 Collection Tubes 75 Elution Tubes 75 Product Insert 1 Documentation PROTOCOLS (63600) Small RNA Library Prep Kit for Illumina - Protocol (6 and 24 prep) SAFETY DATA SHEETS (63600) Small RNA Library Prep Kit for Illumina - Safety Data Sheet Citations Title Metabolite signatures associated with microRNA miR-143-3p serve as drivers of poor lung function trajectories in childhood asthma Journal EBioMedicine. 2024. Authors Kevin M. Mendez, Sofina Begum, Anshul Tiwari, Rinku Sharma, Qingwen Chen, Rachel S. Kelly, et al. Title Circulating miRNAs associate with historical childhood asthma hospitalization in different serum vitamin D groups Journal Respiratory Res. 2024. Authors Hong, X., Jiang, M., Kho, A.T. et al. Title Systems Genomics Reveals microRNA Regulation of ICS Response in Childhood Asthma Journal Cells. 2023. Authors Sharma R, Tiwari A, Kho AT, Celedón JC, Weiss ST, Tantisira KG, McGeachie MJ Title Intrauterine Smoke Exposure, microRNA Expression during Human Lung Development, and Childhood Asthma Journal Int J Mol Sci. 2023. Authors Rosenberg L, Liu C, Sharma R, Wood C, Vyhlidal CA, Gaedigk R, Kho AT, Ziniti JP, Celedón JC, Tantisira KG, et al. Title Obesity and PCOS radically alters the snRNA composition of follicular fluid extracellular vesicles Journal Frontiers in Endocrinology. 2023. Authors Wyse BA, Salehi R, Russell SJ, Sangaralingam M, Jahangiri S, Tsang BK and Librach CL Title Benchmarking omics-based prediction of asthma development in children Journal Respiratory Res. 2023. Authors Wang, XW., Wang, T., Schaub, D.P. et al. Title Characterization of Extracellular Vesicle-Coupled miRNA Profiles in Seminal Plasma of Boars with Divergent Semen Quality Status Journal Int J Mol Sci. 2023. Authors Dlamini NH, Nguyen T, Gad A, Tesfaye D, Liao SF, Willard ST, Ryan PL, Feugang JM. Title Coordinated Regulation of Myonuclear DNA Methylation, mRNA, and miRNA Levels Associates With the Metabolic Response to Rapid Synergist Ablation-Induced Skeletal Muscle Hypertrophy in Female Mice Journal Function (Oxford, England). 2023. Authors Ismaeel A, Thomas NT, McCashland M, Vechetti IJ, Edman S, Lanner JT, Figueiredo VC, Fry CS, McCarthy JJ, Wen Y, Murach KA, von Walden F. Title Obesity and PCOS radically alters the snRNA composition of follicular fluid extracellular vesicles Journal Frontiers in Endocrinology 2023. Authors Wyse, Brandon A., et al. Title Circulating MicroRNAs associated with Bronchodilator Response in Childhood Asthma Journal Research Square (preprint). 2023. Authors Sharma, et al. Title Systems Genomics Reveals microRNA Regulation of ICS Response in Childhood Asthma Journal Cells. 2023. Authors Sharma, R., Tiwari, A., Kho, A. T., Celedón, J. C., Weiss, S. T., Tantisira, K. G., & McGeachie, M. J. Title Expression of Extracellular Vesicle PIWI-Interacting RNAs Throughout hiPSC-Cardiomyocyte Differentiation Journal Frontiers in Physiology. 2022. Authors Louro Ana F., Virgolini Nikolaus, Paiva Marta A., Isidro Inês A., Alves Paula M., Gomes-Alves Patrícia, Serra Margarida. Title Bioactivity and miRNome Profiling of Native Extracellular Vesicles in Human Induced Pluripotent Stem Cell-Cardiomyocyte Differentiation Journal Advanced science (Weinheim, Baden-Württemberg, Germany). 2022. Authors Louro AF, Paiva MA, Oliveira MR, Kasper KA, Alves PM, Gomes-Alves P, Serra M. Title Hydrogel-Embedded Poly(Lactic-co-Glycolic Acid) Microspheres for the Delivery of hMSC-Derived Exosomes to Promote Bioactive Annulus Fibrosus Repair Journal SAGE Journals. 2022. Authors Tyler J., Keti Vaso, and James C. Iatridis, et al. Title Bioactivity and miRNome Profiling of Native Extracellular Vesicles in Human Induced Pluripotent Stem Cell-Cardiomyocyte Differentiation Journal Advanced Science. 2022. Authors Ana F. Louro,Marta A. Paiva,Marta R. Oliveira,Katharina A. Kasper,Paula M. Alves,Patrícia Gomes-Alves,Margarida Serra Title Expression of Extracellular Vesicle PIWI-Interacting RNAs Throughout hiPSC-Cardiomyocyte Differentiation Journal Front Physiol 2022. Authors Ana F. Louro, Nikolaus Virgolini, Marta A. Paiva, Inês A. Isidro, Paula M. Alves, Patrícia Gomes-Alves, and Margarida Serra Title Alteration of miRNAs in Small Neuron‑Derived Extracellular Vesicles of Alzheimer's Disease Patients and the Effect of Extracellular Vesicles on Microglial Immune Responses Journal Journal of Molecular Neuroscience . 2022. Authors Devrim Yagmur Durur, Bora Tastan, Kemal Ugur Tufekci, Melis Olcum, Hamdiye Uzuner, Gökhan Karakülah, Gorsev Yener, Sermin Genc Title Extracellular vesicle characteristics and microRNA content in cerebral palsy and typically developed individuals at rest and in response to aerobic exercise Journal Frontiers in Physiology. 2022. Authors Vechetti IJ, Norrbom J, Alkner B, Hjalmarsson E, Palmcrantz A, Pontén E, Pingel J, von Walden F and Fernandez-Gonzalo R. Title Urine miRNAs as potential biomarkers for systemic reactions induced by exposure to embedded metal Journal Biomarkers in Medicine. 2021. Authors Ivan J Vechetti , Yuan Wen, Jessica F Hoffman, Alexander P Alimov, Vernieda B Vergara, John F Kalinich, Joanna M Gaitens, Stella E Hines, Melissa A McDiarmid, John J McCarthy & Charlotte A Peterson Title High Throughput Sequencing of MicroRNA in Rainbow Trout Plasma, Mucus, and Surrounding Water Following Acute Stress Journal Frontiers in Physiology. 2021. Authors Heather Ikert, Michael D. J. Lynch, Andrew C. Doxey, John P. Giesy, Mark R. Servos, Barbara A. Katzenback, Paul M. Craige Title Distinct Exosomal miRNA Profiles from BALF and Lung Tissue of COPD and IPF Patients Journal International Journal of Molecular Sciences. 2021. Authors Gagandeep Kaur, Krishna Prahlad Maremanda, Michael Campos, Hitendra S. Chand, Feng Li, Nikhil Hirani, M. A. Haseeb, Dongmei Li, Irfan Rahman Title Distinct Exosomal miRNA Profiles from BALF and Lung Tissue of COPD and IPF Patients Journal Int J Mol Sci. 2021. Authors Kaur G, Maremanda KP, Campos M, Chand HS, Li F, Hirani N, Haseeb MA, Li D, Rahman I. Title Circulating MicroRNA: Incident Asthma Prediction and Vitamin D Effect Modification Journal J Pres. Med. 2021. Authors Li J, Tiwari A, Mirzakhani H, Wang AL, Kho AT, McGeachie MJ, Litonjua AA, Weiss ST, Tantisira KG Title Seasonal Variation in miR-328-3p and let-7d-3p Are Associated With Seasonal Allergies and Asthma Symptoms in Children Journal Allergy Asthma Immunol Res.. 2021. Authors Tiwari A, Wang AL, Li J, Lutz SM, Kho AT, Weiss ST, Tantisira KG, McGeachie MJ Title High Throughput Sequencing of MicroRNA in Rainbow Trout Plasma, Mucus, and Surrounding Water Following Acute Stress Journal Frontiers in Physiology. 2021. Authors Ikert H, Lynch MDJ, Doxey AC, Giesy JP, Servos MR, Katzenback BA and Craig PM Title COPD-associated miR-145-5p is downregulated in early-decline FEV1 trajectories in childhood asthma Journal J Allergy Clin Immunol. 2021. Authors Anshul Tiwari, Jiang Li, Alvin T. Kho, Scott T. Weiss, Kelan G. Tantisira, Michael J. McGeachie, et al. Title Circulating MicroRNAs and Treatment Response in Childhood Asthma Journal Am J Respir Crit Care Med. 2020. Authors Jiang Li, Ronald Panganiban, Alvin T. Kho, Michael J. McGeachie, Leanna Farnam, Robert P. Chase, Scott T. Weiss, Quan Lu, and Kelan G. Tantisira Title Exosomal microRNAs are novel circulating biomarkers in cigarette, waterpipe smokers, E-cigarette users and dual smokers Journal BMC Medical genomics. 2020. Authors Singh, K.P., Maremanda, K.P., Li, D. et al. Title Distinctive Cellular Transcriptomic Signature and MicroRNA Cargo of Extracellular Vesicles of Horse Adipose and Endometrial Mesenchymal Stem Cells from the Same Donors Journal Cell Reprogramming. 2020. Authors Felipe Navarrete, Yat Sen Wong, Joel Cabezas, Gonzalo Riadi, José Manríquez, Daniela Rojas, Ana Carolina Furlanetto Mançanares, Lleretny Rodriguez-Alvarez, Fernando Saravia, and Fidel Ovidio Castro. Title MicroRNAs from Extracellular Vesicles Secreted by Bovine Embryos as Early Biomarkers of Developmental Competence Journal Int J Mol Sci. 2020. Authors Melo-Baez B, Wong YS, Aguilera CJ, Cabezas J, Mançanares ACF, Riadi G, Castro FO, Rodriguez-Alvarez L. Title Circulating MicroRNAs and Treatment Response in Childhood Asthma Journal ATS Jounals. 2020. Authors Jiang Li , Ronald Panganiban , Alvin T. Kho , Michael J McGeachie , Leanna Farnam , Robert P. Chase , Scott T Weiss , Quan Lu , and Kelan G Tantisira Title Exosomal microRNAs are novel circulating biomarkers in cigarette, waterpipe smokers, E-cigarette users and dual smokers Journal BMC Medical Genomics. 2020. Authors Kameshwar P. Singh, Krishna P. Maremanda, Dongmei Li & Irfan Rahman Title Serum microRNAs in ASD: Association With Monocyte Cytokine Profiles and Mitochondrial Respiration Journal Frontiers in Psychiatry. 2019. Authors Jyonouchi H, Geng L, Toruner GA., Rose S, Bennuri SC. and Frye RE Title Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers Journal J Extracellular Vesicles. 2019. Authors Isaac Kirubakaran Sundar, Dongmei Li, Irfan Rahman Title Extracellular vesicles from endometriosis patients are characterized by a unique miRNA-lncRNA signature Journal JCI Insight. 2019. Authors Kasra Khalaj, Jessica E. Miller, Harshavardhan Lingegowda, Asgerally T. Fazleabas, Steven L. Young, Bruce A. Lessey, Madhuri Koti, and Chandrakant Tayade Title Comprehensive Profiling of the Circulatory miRNAome Response to a High Protein Diet in Elderly Men: A Potential Role in Inflammatory Response Modulation. Journal Mol Nutr Food Res. 2019 Apr;63(8):e1800811. Authors Ramzan F, Mitchell CJ, Milan AM, Schierding W, Zeng N, Sharma P, Mitchell SM, D'Souza RF, Knowles SO, Roy NC, Sjödin A, Wagner KH, Cameron-Smith D. Title Extracellular vesicles from endometriosis patients are characterized by a unique miRNA-lncRNA signature Journal JCI Insight. 2019. Authors Kasra Khalaj, Jessica E Miller, Harshavardhan Lingegowda, Asgerally T Fazleabas, Steven L Young, Bruce A Lessey, Madhuri Koti, Chandrakant Tayade Title Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers Journal Journal of Extracellular Vesicles, 8(1). 2019. Authors Sundar, I. K., Li, D., & Rahman, I. Title Diversity and signature of small RNA in different bodily fluids using next generation sequencing Journal BMC genomics, 19(1), 408. 2018. Authors El-Mogy, M., Lam, B., Haj-Ahmad, T. A., McGowan, S., Yu, D., Nosal, L., ... & Haj-Ahmad, Y. Title Diversity and signature of small RNA in different bodily fluids using next generation sequencing Journal BMC Genetics. 2018. Authors El-Mogy, M., Lam, B., Haj-Ahmad, T.A. et al. Related Services NGS Related Products Total RNA Purification Kit Plasma/Serum RNA Purification Kit Low Abundance RNA Quantification Kit
Small RNA Library Prep Kit for Illumina All the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Small RNA Library Prep Kit for Illumina All the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform Price USD$1,635.00 SKU 63600 Format Indexes 1-24 Indexes 25-48 Size 24 Preps Quantity