Prokaryotic RNA Sequencing
Service Specification and Delivery
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| rRNA depleted directional library | Illumina PE150 | 1-2 Gb | Q30≥85% |
Sample Requirement
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Limited or no protein or DNA contamination shown on gel. |
RIN≥6.5 |
Eukaryotic mRNA Sequencing
Service Specifications
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| Poly A enriched | Illumina PE150 DNBSEQ-T7 |
6-10 Gb | Q30≥85% |
Minimum Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity | |
|---|---|---|---|---|
| Standard Library | ≥ 10 | ≥ 0.2 | 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≥4.0 For animals: RIN≥4.5; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Directional Library | ≥ 10 | ≥ 0.2 | 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≥4.0 For animals: RIN≥4.5; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 300 mg Viscera or Brain: 240 mg Muscle: 450 mg Bones, Hair or Skin: 1 g |
Insects: 6 g Crustacea: 300 mg |
1 tube | 106 cells |
Non-Reference Based mRNA Sequencing
Service Specifications
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| Poly A enriched | Illumina PE150 DNBSEQ-T7 |
6-10 Gb | Q30≥85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 10 | ≥ 0.2 | 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≥4.0 For animals: RIN≥4.5; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 300 mg Viscera or Brain: 240 mg Muscle: 450 mg Bones, Hair or Skin: 1 g |
Insects: 6 g Crustacea: 300 mg |
1 tube | 106 cells |
Full-Length mRNA Sequencing - Nanopore
Service Specifications
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| Poly A enriched | Nanopore PromethION 48 | 6/12 Gb | Average quality score: Q10 |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 100 | ≥ 1.0 | 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≥7.0 For animals: RIN≥7.5; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 300 mg Viscera or Brain: 240 mg Muscle: 450 mg Bones, Hair or Skin: 1 g |
Insects: 6 g Crustacea: 300 mg |
1 tube | 106 cells |
Full-Length mRNA Sequencing - PacBio
Service Specifications
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| Poly A enriched mRNA CCS library | PacBio Sequel II PacBio Revio |
20/40 Gb 5/10 M CCS |
Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 100 | ≥ 1.0 | 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≥8.0; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 300 mg Viscera or Brain: 240 mg Muscle: 450 mg Bones, Hair or Skin: 1 g |
Insects: 6 g Crustacea: 300 mg |
1 tube | 106 cells |
PacBio 2+3 Full-Length mRNA Solution
Service Specifications
| Library | Sequencing strategy | Data recommended | Quality Control |
|---|---|---|---|
| Poly A enriched mRNA CCS library | PacBio Sequel II PacBio Revio |
20/40 Gb 5/10 M CCS |
Q30 ≥ 85% |
| Poly A enriched | Illumina PE150 | 6-10 Gb | Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity | |
|---|---|---|---|---|
| Illumina Library | ≥ 10 | ≥ 0.2 | 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≥4.0 For animals: RIN≥4.5; 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| PacBio Library | ≥ 100 | ≥ 1.0 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
Plants: RIN≥7.5 Animals: RIN≥8.0 5.0≥28S/18S≥1.0; limited or no baseline elevation |
Long Non-Coding RNA Sequencing - Illumina
Service Specifications
| Library | Platform | Data recommended | Quality Control |
|---|---|---|---|
| rRNA depleted directional Library | Illumina PE150 | 10-16 Gb | Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 80 | ≥ 0.8 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
RIN≥6.0 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 450 mg Viscera or Brain: 240 mg Muscle: 600 mg Bones, Hair or Skin: 1.5 g |
Insects: 9 g Crustacea: 450 mg |
2 tube | 106 cells |
Small RNA Sequencing - Illumina
Service Specifications
| Library | Platform | Data recommended | Quality Control |
|---|---|---|---|
| Size selected | Illumina SE150 | 10M-20M reads | Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 80 | ≥ 0.8 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
RIN≥6.0 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 450 mg Viscera or Brain: 240 mg Muscle: 600 mg Bones, Hair or Skin: 1.5 g |
Insects: 9 g Crustacea: 450 mg |
2 tube | 106 cells |
CircRNA Sequencing - Illumina
Service Specifications
| Library | Platform | Data recommended | Quality Control |
|---|---|---|---|
| rRNA depleted directional Library | Illumina PE150 | 16-20 Gb | Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 80 | ≥ 0.8 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
RIN≥6.0 5.0≥28S/18S≥1.0; limited or no baseline elevation |
| Plants | Animal | Arthropods | Whole blood | Cells |
|---|---|---|---|---|
| Root, Stem or Petal: 450 mg Leaf or Seed: 300 mg Fruit: 1.2 g |
Heart or Intestine: 450 mg Viscera or Brain: 240 mg Muscle: 600 mg Bones, Hair or Skin: 1.5 g |
Insects: 9 g Crustacea: 450 mg |
2 tube | 106 cells |
Exosomal Small RNA Sequencing - Illumina
Service Specifications
| Library | Platform | Recommended data | Data QC |
|---|---|---|---|
| Exosomal miRNA isolation | Illumina SE50 | 10M-20M reads | Q30 ≥ 85% |
Sample Requirement
| Amount (ng) | Volume (μL) |
|---|---|
| 40 | 20 |
|
Exosome: 100 μL; 40 μg; 109 particles/mL Serum/Plasma: 5 mL, Cell culture supernatant: 100 mL, Urine: 100 mL, Saliva: 15 mL, Nasal discharge: 15 mL, Cerebrospinal: 15 mL, Amniotic fluid: 50 mL, Semen: 5 mL Bile: 5 mL, Ascites: 50 mL, Milk: 15 mL, Follicular fluid: 15 mL, Fecal supernatant: 50 mL, Microbial culture supernatant: 15 mL Animal tissue: 3 g, Plant tissue: 1 g |
|
Exosomal mRNA/LncRNA/CircRNA Sequencing - Illumina
Service Specifications
| Library | Platform | Recommended data | Data QC |
|---|---|---|---|
| Exosomal miRNA isolation | Illumina PE150 | 10-16 Gb | Q30 ≥ 85% |
Sample Requirement
| Amount (ng) | Volume (μL) |
|---|---|
| 40 | 20 |
|
Exosome: 100 μL; 40 μg; 109 particles/mL Serum/Plasma: 5 mL, Cell culture supernatant: 100 mL, Urine: 100 mL, Saliva: 15 mL, Nasal discharge: 15 mL, Cerebrospinal: 15 mL, Amniotic fluid: 50 mL, Semen: 5 mL Bile: 5 mL, Ascites: 50 mL, Milk: 15 mL, Follicular fluid: 15 mL, Fecal supernatant: 50 mL, Microbial culture supernatant: 15 mL Animal tissue: 3 g, Plant tissue: 1 g |
|
Metatranscriptome Sequencing
Service Specifications
| Sequencing platform | Sequencing Strategy | Data recommended | Data Quality Control |
|---|---|---|---|
| Illumina NovaSeq | PE150 | 12 Gb | Q30 ≥ 85% |
Sample Requirements
| Concentration (ng/μL) | Total amount (μg) | Volume (μL) | OD260/280 | OD260/230 | RIN |
|---|---|---|---|---|---|
| ≥ 50 | ≥ 1.0 | ≥ 20 | 1.8-2.0 | 1.0-2.5 | ≥ 6.5 |
Whole Transcriptome Sequencing - Illumina
Service Specifications
| Library | Sequencing Strategy | Data recommended | Quality Control |
|---|---|---|---|
| rRNA depleted | Illumina PE150 | 16-20 Gb | Q30 ≥ 85% |
| Size selected | Illumina SE50 | 10-20M reads | Q30 ≥ 85% |
Sample Requirements
| Conc. (ng/μl) | Amount (μg) | Purity | Integrity |
|---|---|---|---|
| ≥ 80 | ≥ 1.6 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Limited or no protein or DNA contamination shown on gel. |
RIN ≥ 6.0 5.0 ≥ 28S/18S ≥ 1.0; limited or no baseline elevation |
Spatial Transcriptome
1. BMKMANU S3000 Spatial Transcriptome
| Sample Requirements | Library | Sequencing Strategy | Data Recommended | Quality Control |
|---|---|---|---|---|
| OCT-embedded cryo samples (approx. 6×6×6 mm³) 2 blocks per sample (1 for experiment, 1 for backup) |
S3000 cDNA Library | Illumina PE150 | 160K PE reads per 100uM (250 Gb) | RIN > 7 |
2. 10x Genomics Visium Spatial Transcriptome
| Sample Requirements | Library | Sequencing Strategy | Data Recommended | Quality Control |
|---|---|---|---|---|
| OCT-embedded cryo samples (approx. 6×6×6 mm³) 2 blocks per sample |
10X Visium cDNA Library | Illumina PE150 | 50K PE reads per spot (60 Gb) | RIN > 7 |
Single-Nuclei RNA Sequencing
Samples eligible for single nuclei RNA sequencing
| Cell / Tissue | Reason |
|---|---|
| Unfresh frozen tissue | Unable to get fresh or long-saved organizations |
| Muscle cell, Megakaryocyte, Fat... | Cell diameter is too large to enter the instrument |
| Liver... | Too fragile to break, unable to distinguish single cells |
| Neuron cell, Brain... | More sensitive, easy to stress, will change the sequencing results |
| Pancreas, Thyroid... | Rich in endogenous enzymes, affecting the production of single cell suspension |
Single-nucleus vs Single-cell
| Single-nucleus | Single-cell |
|---|---|
| Unlimited cell diameter | Cell diameter: 10-40 μm |
| The material can be frozen tissue | The material must be fresh tissue |
| Low stress of frozen cells | Enzyme treatment may cause cell stress reaction |
| No red blood cells need to be removed | Red blood cells need to be removed |
| Nuclear expresses bioinformation | The whole cell expresses bioinformation |
Service Specifications
| Sample Requirements | Library | Sequencing Strategy | Data Recommended | Quality Control |
|---|---|---|---|---|
| Animal tissue ≥ 200 mg Plant tissue ≥ 400 mg |
10x Genomics sn cDNA library | Illumina PE150 | 100K PE reads per cell (100-200 Gb) | 700-1200 nuclei/μl and nuclei integrity observed under microscope |