Total RNA sequencing captures both coding and non-coding RNAs by removing rRNA instead of enriching poly(A) RNA. This approach enables comprehensive transcriptome analysis, including mRNA, lncRNA, circRNA, and other regulatory transcripts.
This makes Total RNA-Seq increasingly important in systems biology, disease research, and transcriptome discovery.
Major Application Areas
Total RNA library preparation is widely used for:
|
Application |
Description |
|
Whole transcriptome profiling |
Analyze coding and non-coding RNA |
|
Non-coding RNA discovery |
Identify novel regulatory RNAs |
|
Host–pathogen studies |
Explore infection-related transcript changes |
|
Degraded sample sequencing |
Suitable for FFPE RNA samples |
Because it retains a broader RNA population, this approach is particularly valuable for exploratory transcriptomics.
Industry Pain Points
Researchers performing Total RNA-Seq often face challenges such as:
|
Challenge |
Impact |
|
High rRNA background |
Wastes sequencing reads |
|
Complex depletion protocols |
Increases workflow time |
|
Low-abundance transcripts |
Reduced detection sensitivity |
Since rRNA accounts for 80–90% of total RNA, efficient removal is essential.
Yeasen Total RNA Library Preparation Workflow
Yeasen provides comprehensive total RNA library preparation workflows, offering both strand-specific and non-strand-specific library construction options in a single system. In addition, the premixed total RNA library preparation kit simplifies experimental steps and streamlines the workflow, enabling fast and efficient library preparation.
Typical workflow includes:
The optimized protocol improves informative sequencing reads and transcript coverage.
Key Experimental Tips
|
Factor |
Recommendation |
|
rRNA removal |
Ensure high depletion efficiency |
|
RNA input |
Use sufficient starting material |
|
RNA integrity |
Minimize degradation |
|
PCR cycles |
Optimize amplification steps |
When properly optimized, Total RNA-Seq provides a comprehensive view of transcriptome dynamics.
FAQ
Q1. Why is lncRNA-seq different from mRNA-seq?
Many long non-coding RNAs lack poly(A) tails, making poly(A) enrichment insufficient. lncRNA sequencing usually relies on rRNA depletion instead of poly(A) selection.
Q2. What types of transcripts can be detected with lncRNA-seq?
lncRNA-seq enables detection of both polyadenylated and non-polyadenylated transcripts, including regulatory lncRNAs, antisense RNAs, and intergenic transcripts.
Q3. Is RNA quality critical for lncRNA sequencing?
lncRNA-seq is generally more tolerant of partially degraded RNA because rRNA depletion does not depend on intact poly(A) tails.
Q4. What research applications benefit from lncRNA-seq?
Common applications include epigenetic regulation studies, cancer transcriptomics, RNA regulatory networks, and functional annotation of non-coding RNAs.
Related Product
|
Category |
Name |
Cat. No. |
Size |
|
|
RNA Lib Prep |
Dual-mode(Strand specific & Non Strand specific) |
12308ES24/96 |
24 T/96 T |
|
|
Premix version |
12340ES24/96 |
|||
|
12341ES24/96 |
||||
|
mRNA isolation |
Eukaryotic mRNA |
12629ES24/96 |
24 T/96 T |
|
|
rRNA depletion |
Human/Mouse/Rat |
Hieff NGS™ MaxUp Human/Mouse/Rat rRNA Depletion Kit(rRNA ITS/ETS) |
12257ES24/96 |
|
|
Hieff NGS™ MaxUp Human/Mouse/Rat rRNA Depletion Kit(rRNA ITS/ETS) 2.0 |
12726ES24/96 |
|||
|
Plant |
12254ES24/96 |
|||
|
Beads |
- |
12602ES03/08/56 |
1/5/60 mL |
|