The NEBNext Small RNA library prep is a commercially available kit from New England Biolabs that provides a novel protocol that results in higher yields and lower adapter-dimer contamination. The protocol generates small RNA libraries directly from total RNA. MicroRNAs (miRNAs) having undergone processing by the Dicer complex are efficiently targeted by adapter ligation to the 5' phosphate and 3' OH modifications of the miRNAs.
Indexes are available for multiplexing of up to 48 samples. Total RNA input is 100 ng - 1000 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 small RNA 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 |
| 100-1000 ng | 50 - 150 ng/μl 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 | ||
| 10-20 million reads per sample | Single end | Single index | 75 bases | ||
* minimum read count recommended by MUGTC; increased sequencing depth may be desired for some studies