● Capture of poly-A mRNA followed by cDNA synthesis and library preparation
● Sequencing of the full-length transcripts
● Bioinformatic analysis based on alignment to a reference genome
● Bioinformatic analysis includes not only expression at gene and isoform-level but also analysis of lncRNA, gene fusions, poly-adenylation and gene structure
|
Library |
Sequencing strategy |
Data recommended |
Quality Control |
|
Poly A enriched |
Illumina PE150 |
6 Gb |
Average quality score: Q10 |
|
Conc.(ng/μl) |
Amount (μg) |
Purity |
Integrity |
|
≥ 100 |
≥ 0.6 |
OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
For plants: RIN≥7.5; For animals: RIN≥7.0; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
● Plants:
Root, Stem or Petal: 450 mg
Leaf or Seed: 300 mg
Fruit: 1.2 g
● Animal:
HEart or Intestine: 300 mg
Viscera or Brain: 240 mg
Muscle: 450 mg
Bones, Hair or Skin: 1g
● Arthropods:
Insects: 6g
Crustacea: 300 mg
● Whole blood: 1 tube
● Cells: 106 cells
Container: 2 ml centrifuge tube (Tin foil is not recommended)
Sample labeling: Group+replicate e.g. A1, A2, A3; B1, B2, B3.
Shipment:
1. Dry-ice: Samples need to be packed in bags and buried in dry-ice.
2. RNAstable tubes: RNA samples can be dried in RNA stabilization tube(e.g. RNAstable®) and shipped in room temperature.