mRNA Enrichment

PolyA enrichment library preparation positively selects for transcripts with polyadenylated tails and is often referred to as mRNA sequencing. All mRNA and noncoding transcripts with polyA tails are captured via hybridization to polyT oligos on magnetic beads, leaving untailed molecules behind. RNA integrity is of particular importance with this method as degradation of the polyA tail will result in reduced mRNA capture and may affect downstream analysis. As only polyA tailed transcripts will be included in the final dataset, this is the first choice for many eukaryotic studies looking for changes in coding regions.

For this pipeline, SeqCenter uses the Illumina Stranded mRNA Prep kit to generate unbiased RNA libraries. mRNA enrichment is a more cost-effective method and generates greater exonic coverage; however, the major limitation is that it can only be used for organisms with polyA tails. It also requires high-quality RNA and higher minimum sample concentrations.

We strongly recommend visiting our overall Illumina RNA services page for additional considerations when planning any RNA experiment, and we encourage all customers to check out the tips for handling RNA and guidance when assessing sample quality for submission.

Sample Requirements:

  • Samples shipped frozen on dry ice
  • Total volume of 20 µL or more
  • Sample concentration greater than 50 ng/uL
  • DNAse treated (Recommended, not required)
  • RIN > 6 (Recommended measurement, not required)

What You Will Receive:

  • Two fastq files (2 x 150bp) for each sample
  • Sequencing stats summary
  • Materials and methods summary
  • Data shared via Box folder within 2 weeks

Packages & Pricing

Sequencing Package
(Minimum Read Count Per Sample)
25M Paired End Reads (3.75 Gbp)
Example uses: Yeast
50M Paired End Reads (7.5 Gbp)
Example uses: Larger fungi, fruit flies, algae
100M Paired End Reads (15 Gbp)
Example uses: Mouse and human
200M Paired End Reads (30 Gbp)
Example uses: Transcriptome assembly
*Orders >48 Samples receive 5% discount

We offer two levels of RNA analysis packages, Basic and Intermediate, which can help our customers to start analyzing their differential expression studies. These packages are a great starting place for burgeoning bioinformaticians, but they will not pick up non-coding RNA and are not intended for metatranscriptomes. For more information, please visit our RNA Analysis services page.

Due to the sensitive nature of RNA and RNA-seq projects, we recommend an approach tailored to a project’s organism and experimental setup that is carried out in a uniform manner soon after harvesting.

For customer convenience, we do offer RNA extraction services using the Zymo Quick-RNA Miniprep Kit, which is broadly applicable but may not be optimal for all sample types or projects. This kit enables us to quickly isolate DNA-free RNA from cell pellets as well as homogenized tissue samples. We recommend sending cell pellets that contain at least 107-108 cells, in which the pellet is sizable and clearly visible in the tube. For tissue samples, we ask that you homogenize them as much as possible to enable successful extractions.

This extraction kit is compatible with ZymoResearch’s DNA/RNA Shield, and it should be noted on orders when samples have been treated with this product. More info on the Quick-RNA Miniprep Kit can be found here.

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