RNA-seq analysis
Overview
This service is for RNA sequencing analysis in which total RNA is to be submitted, libraries are prepared using the Biomek i7 automated workstation, sequencing is performed, and fastq data is delivered.
We use the following kits from New England Biolabs for library preparation.
- NEBNext Poly(A) mRNA Magnetic Isolation Module[E7490]
- NEBNext Ultra II Directional RNA Library Prep Kit [E7760]
- NEBNext Multiplex Oligos for Illumina (96 Unique Dual Index Primer Pairs) [E6440]
Sample Submission Guidelines
Total RNA
- Volume: minimum 8 µl
- Total amount: minimum 300 ng
- Concentration: 10-150ng/µl, recommended 50–100 ng/µl
- We recommend DNase treatment during the RNA purification process. (Not required).
- Measure the RNA concentration yourself (e.g., Qubit, Nanodrop).
- 8-strip tubes can be used for 8 or more samples, or 96-well plates for 24 or more samples.
- SignAC does not measure RNA integrity prior to library preparation. It is recommended that the library preparation kit used has a RIN value of 7 or higher. We recommend that you check this using Agilent Bioanalyzer, etc. (Any problems caused by RNA malfunctions are the responsibility of the users.)
- Please submit an Excel file with the sample name (information written on the tube) and concentration through the Redmine system.
- Add a serial number to the sample name and write that number to the tube as well.
- As a general rule, proceed with library preparation based on the concentration specified by each individual.
- Please write down your name and the Redmine ticket number down on your sample tube.
Data Delivery
fastq files
- Sequencing is targeted at 50 bp paired end and over 15 million reads per sample.
- Adapter trimming is not performed.
- Please contact us for a simple primary analysis (e.g., mapping, etc.).
Estimated delivery date
Please contact us.
Reference
1 month (FY2022 results).