RNA starting material requirements

Sample preparation options           RNA Amount         Minimal concentration    Minimal volume
INCPM mRNASeq 100-500 ng 20 ng/μl 5 μl
INCPM mRNASeq w/o polyA selection*      80 ng 13 ng/μl 6 μl
Bulk MARS-seq 1-50 ng 5 ng/μl ** 6.1 μl
Illumina TruSeq Stranded Total RNA 0.1 - 1 μg   5-10 μl
Illumina TruSeq Stranded mRNA 0.1 - 4 μg   5-50 μl
Illumina TruSeq Small RNA 1 μg 200 ng/μl  
Clontech SMARTer v4 Ultra Low RNA 10 pg - 10 ng Up to 9.5 μl  
Bulk SMART-seq 2 0.1-50 ng   3.2 μl

 

* For RNA samples that do not contain polyA, please perform ribosomal depletion or contact us.

** For submitting RNA for bulkMARS-seq see guidlines here.

  • Please submit twice the amount from that noted in the table
  • Please add extra volume to enable accurate pippettation
  • Measure RNA conenctration by nanodrop or Qubit (do not rely on Bioanalyser or Tapestation concentration).

RNA submission criteria

Quality parameters RNA
OD260/280 ≥ 1.8
OD260/230 ≥ 2
RIN number ≥ 8
  • Please note that Agilent RIN numbers are based on human, mouse, and rat RNA profiles. Other organisms’ profiles may not generate high RIN numbers, but may still be high quality. You should find the expected profile for your species and assess the quality.
  • RMA samples that underwent polyA selection or ribosomal depletion cannot be evaluated for RIN score (as it is based on ribosomal RNA)
  • Note that the user is responsible for the results in case the sample does not match the specifications above, or do not meet the criteria of the procedure requested.