Table 2 Proposed guidelines for both wet- and dry-lab criteria. List of the important criteria at each step of the workflow and the corresponding recommended thresholds based on our study

Extraction

RNA quantity

100 ng

Library preparation

RNA molarity

 > 5 nM

Sequencing preprocessing

Sequencing depth

 > 50 M reads

GC content

Unique peak

Mapping

rRNA after depletion

 < 10%

Globin after depletion

 < 10%

Library strandness (% undesirable sens)

 < 10%

Genome mapping

 > 80%

Protein coding content

 ~ 75%

Quantification

Rarefaction curves

Reach a plateau with homogeneity

Number of detected genes

Human: [12500-14000]

NHP: [12500–14000]

Mouse: [10500–12000]

Rabbit: [10500–12000]