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RNA Sequencing (RNA-Seq)


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RNA-Seq, also known as Whole Transcriptome Shotgun Sequencing (WTSS), is a revolutionary high-throughput sequencing method for the evaluation of RNA quantities (Wang, Gerstein et al. 2009). Usually performed in combination with next-generation sequencing (NGS), the RNA-Seq technique allows the determination of gene transcriptional structures, splicing patterns, post transcriptional modifications, expression levels, and so on.

A regular RNA-Seq assay generally requires three steps (Figure 1):

  1. RNA library preparation
    1. Total RNA extraction
    2. RNA fractionation and selection: for differential experimental purposes, specific fractions of RNAs are isolated for subsequent analysis.
    3. cDNA construction via reverse transcription
  2. High-throughput deep sequencing
  3. Computational data analysis

RNA-Seq-pic1.jpg
Figure 1. A description of the workflow for a regular RNA-Seq assay (Wang, Gerstein et al. 2009).

Since its development, the RNA-Seq technique has gained tremendous popularity, due to its innate advantages over other methods, including:

  • High-throughput capacity
  • High resolution (single-base resolution)
  • High level of reproducibility
  • Low background noise
  • Potential to reveal sequence variations such as SNPs
  • Feasibility for non-model organisms with no existing genomic sequence
  • Requires lower amount of RNA
  • Relatively low cost

In the field of plant biology, the RNA-Seq technique has also been adopted for the study of a wide range of scientific topics, such as profiling of temporal, special and evolutional transcriptome landscapes, identification and characterizing of novel non-coding RNAs, gene function analysis, establishment of gene co-expression networks.

Lifeasible is a leading plant biotechnology company with proven expertise in plant genetics and plant molecular biology. We proudly provide one-stop plant RNA-Seq service from experimental design to data presentation. Our services are featured by:

  • Customized sample preparation: we provide optimized protocols for the handling and RNA extraction of different types of plant samples, including plant cells, tissues, organs as well as whole plant.
  • Multiple options for RNA preparation: with the adjustability of our service to various research goals, differential RNA fractions, including messenger RNA (mRNA), microRNA (miRNA), long non-coding RNA (LncRNA) and circular RNA (circRNA) can be obtained with high quality.
  • Cutting-edge next-generation sequencing: our high-capacity sequencing facilities guarantee maximum output of your experiment, by providing clean data with high confidence.
  • Comprehensive data analysis. Our groups of scientists, bioinformaticians and computational specialists always work closely with our customers for data analysis, interpretation, organization and presentation.

Welcome to contact us for questions, inquiries or collaborations.

Reference

  1. Wang, Z., M. Gerstein and M. Snyder (2009). "RNA-Seq: a revolutionary tool for transcriptomics." Nat Rev Genet 10(1): 57-63.
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