The SMART-Seq v4 Ultra Low Input RNA Prep is a commercially available kit from Takara Bio employed to construct RNA-Seq libraries from total RNA of <10 ng. SMART-Seq technology uses oligo(dT) priming and proprietary template switching technology to generate full-length cDNA from intact full-length mRNA. Libraries are constructed from the cDNA samples with reagents supplied in the NEBNext Ultra II FS DNA Library Prep Kit. Total RNA integrity is evaluated for each sample prior to library construction by processing on a Fragment Analyser to view the rRNA profile.
Unique dual-indexes are available for multiplexing of up to 96 samples. Total RNA input is 10 pg - 10 ng.
Core staff are available to assist with experimental design and to answer questions related to next-generation sequencing technology. Arrangements for a meeting to discuss new or existing projects can be made by sending an email to Nathan Bivens, Genomics Core director.
All projects must be initiated with a quote before samples are submitted. Send a quote request to the general core email mugenomicscore@missouri.edu.
The RNA-Seq Calculator may be used to estimate the amount of sequence required for a project.
General Considerations:
- Submit samples only after paperwork has been completed.
- Clearly label tubes. Names must match those given on the project information form.
- Low binding microcentrifuge tubes (0.5 or 1.5 mL) are preferred for storing high quality nucleic acids.
Submission Requirements:
| Input Range | Submission Concentration |
Recommended Buffer |
Special Instructions |
| 10 pg - 10 ng | 1 - 10μl total RNA | nuclease free water | Provide sample source and extraction methods used. DNase treatment strongly recommended. |
| Sequence Depth/Coverage* | Read Type | Index Type | Read Length | ||
| 50 million reads per sample | Paired end | unique dual-index | 100 bases | ||
* minimum read count recommended by MUGTC; increased sequencing depth may be desired for some studies