NGS for Small RNA and microRNA

Norgen Biotek offers comprehensive services for Next-generation sequencing (NGS) in an accredited state-of-the-art laboratory from sample isolation to sequencing. Norgen Biotek has extensive expertise in sample preparation, sequencing and analysis of all types of samples: cells, tissues, exosomes, as well as bodily fluids including plasma, serum, urine, saliva, etc, bacteria, viruses, and plant tissues. Our expert staff will assist in the initial consultation and sample handling to the final analysis of data presented in an easy to understand report. Your project will receive full attention by experts with a guaranteed fast turn-around time. Our services include: 1) Sample Isolation, 2) RNA Quanitification and QC, 3) Library Preparation, 3) Library Quantification and QC, 4) Sequencing, and 5) Data Analysis, 6) Data Report.

 

  • Comprehensive services - sample-to-answer
  • Expertise in RNA extraction and sequencing of all types of samples including bodily fluid samples
  • Excellent ability to derive meaningful data from samples with low RNA concentration
  • Close consultation and expert advice throughout the process
  • Quality assessment of RNA at every step of the process
  • Fast turn-around and very responsive staff
  • Accredited lab with the highest standard of proficiency

Service Workflow:

  1. Consultation - This is free of charge. It is essential to have an initial engagement regarding the samples and the sequencing project with our experts to define and set clear objectives and outcomes. This is an excellent way to start your project. What are you trying to achieve? Depth of reads required to answer your questions, etc.?
  2. Sample Submission - We will provide instructions on how to handle, process, and ship the sample and specify the sample requirement (based on sample type). In addition, we will offer sample isolation services.
  3. Sample Isolation - We have tremendous expertise for all sample extractions including low concentration RNA samples from bodily fluids and other sample types. We offer high quality kits for sample isolation or we can do the sample isolation at our lab with scientists that have expertise in the sample types.
  4. RNA Quality Control (QC) - We perform quality control on the isolated RNA to assess quantity, quality, and amplifiability prior to library prep. We share this data prior to proceeding with sequencing.
  5. Library Preparation - We generate library preps using industry standard kits for generating small RNA libraries.
  6. Sequencing - We use the Illumina platform for sequencing microRNA for profiling and discovery.
  7. Analysis - Complete Bioinformatics analysis.
    • Mapping/alignment of all quality reads to reference database(s): miRBase, species genome, etc., classification, and functional annotation of all mapped reads.
    • Biostatistical analysis: Expression analysis, multi-parameter data analysis, length distribution, transcript copy number comparisons, etc., length distribution of counts of sRNAs, conservation profile of the identified miRNAs analysis, differential expressed analysis, pre-miRNA clusters, miRNA target prediction, miRNA target GO and KEGG enrichment analysis (optional).
  8. Report - A comprehensive report is the final deliverable and is submitted in a timely fashion. All data can be analyzed and raw data can be downloaded from a secure site.

 

Small RNA Sequencing from as little as 50 μL of Plasma. Norgen Biotek has developed an effective pipeline of small RNA sequencing from small volume of plasma or serum. RNA could be effectively and consistently recovered from as little as 50 μL of plasma using Norgen’s patented sample preparation technology (example shown here with Plasma/Serum RNA Purification Mini Kit, Cat# 55000). Panel A showed that the number of microRNA detected from 50 or 200 μL of Plasma was almost identical to that of 4 mL. Panel B is a Venn diagram showing that of the microRNAs identified, the majority are detected in all volume of plasma input from 50 μL to 4 mL. In fact, the scatter plots in Panel C showed the relative expression level of each microRNA detected was highly correlated between 50 or 200 μL of Plasma and 4 mL plasma.

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Norgen Biotek RNA Sequencing Process Workflow:

Initial Consultation

Expert advice on sample handling, processing and experimental design for best data outcome

RNA Isolation/RNA Submission

You can elect to simply submit isolated RNA or send the samples for RNA isolation; our team will guide you through what is required from different sample types

RNA Quality Control

Isolated RNA will be quantified, assessed for quality and PCR amplification

Library Preparation

Library preparation with industry standard kits, QC and quantification, Size selection

Sequencing

Illumina platform

Data Analysis

Filtering, aligning/mapping, annotating with statistical analysis

Final Report

Comprehensive report detailing the analysis will be the final deliverable