New RNA sequencing methods enable discovery and profiling of RNAs in any organism without prior genome annotation. Rapid advancement of new technologies has led to the development of several methods of RNA sequencing which differ in many respects such as: the RNA selection method, required sequencing depth, and type of data generated. The purpose of this video is to explain the differences between the various RNA sequencing services that LC Sciences offers, and help you determine which service is right for your project based on the goals of your experiment(s).
| Description | Poly(A) RNA-Seq | Single-Cell RNA-Seq |
Total RNA-Seq | microRNA-Seq | Degradome Sequencing | lncRNA-Seq | circRNA-Seq | circRNA-Seq with ID | Whole Transcriptome Sequencing |
| RNA Species | mRNA | mRNA | mRNA | microRNA | mRNA fragments | mRNA + lncRNA | mRNA + lncRNA + circRNA | circRNA | mRNA, miRNA, lncRNA, circRNA |
| RNA Selection Method | PolyA-Tail Selection | PolyA-Tail Selection | rRNA Depletion | Size Fractionation | PolyA-Tail Selection – mRNA fragments | rRNA Depletion | rRNA Depletion | rRNA Depletion & RNase R Digest | rRNA Depletion & Size Fractionation |
| Data Analysis | Discovery, Expression, Annotation | Expression, Annotation | Discovery, Expression, Annotation | Discovery, Expression, Annotation | miRNA Target ID, Degradation Plots | Discovery, Expression, Annotation | Discovery, Expression, Annotation | Discovery, Expression, Annotation | Discovery, Expression, Annotation |
| Sequencing Instrument | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 | HiSeq 4000 |
| Sequencing Run Length | 150bp Paired-end | 150bp Paired-end | 150bp Paired-end | 50bp Single-end | 50bp Single-end | 150bp Paired-end | 150bp Paired-end | 150bp Paired-end | 150bp Paired-end & 50bp Single-end |
| Recommended Sequencing Depth | 40 million | 100k reads/cell (may vary) | 40 million | 7-10 million | 7-10 million | 80 million | 80 million | 40 million | 80 million & 7-10 million |
| Type of Sample | Total RNA | Viable Cells | Total RNA | Total RNA | Total RNA | Total RNA | Total RNA | Total RNA | Total RNA |
| Pref Sample Amount | 4 µg | N/A | 4 µg | 1 µg | 10 µg | 4 µg | 4 µg | 4 µg | 4 µg |
| Min Sample Amount Required | 2 µg | N/A | 2 µg | 100 ng | 5 µg | 2 µg | 2 µg | 2 µg | 2 µg |
* Inquire about immuno-precipitated RNAs or other enriched RNAs.