Sample Requirements

DNA Sample Requirements

RNA Sample Requirements

Proteomics Sample Requirements

FFPE Sample Requirements

 

DNA

DNA Sample Requirements

Please provide sample analysis results measured by Qubit, NanoDrop, AGE or Agilent 2100 in one or multiple forms. For large inserted sized DNA library, the samples should be based on Qubit or AGE quantification. NanoDrop quantification is not recommended.

de novo Sequencing:

Sample quantity required (single pair):

  1. Short-insert libraries: ≥2.5 µg
  2. 2 kb large-insert libraries: ≥20 µg
  3. 5 kb-6 kb large-insert libraries: ≥20 µg
  4. 10 kb large-insert libraries: ≥30 µg
  5. 20 kb and 40 kb large-insert libraries: ≥60 µg
  6. PCR-free libraries with high or low GC content: ≥30 µg

Note: the total sample quantity required is also determined by the experimental strategy, as well as the type and number of libraries to be constructed.

Sample concentration:

  1. Short-insert libraries: ≥25 ng/ µL
  2. Large-insert libraries: ≥133 ng/ µL

Sample quality: genomic DNA should be intact.
Sample purity:
OD260/280= 1.8-2.0

Whole Genome Resequencing:

Plants and Animals Whole Genome Resequencing:

For the genomic DNA samples you will provide:
Purity: OD 260/280=1.8~2.2
Total DNA: Sample quantity demanded: ≥2.5μg/library
DNA Concentration: ≥25ng/μl, the greater the amount the better

Microbial Whole Genome Resequencing:

For the genomic DNA samples you provide us:
Purity: OD260/280=1.8-2.0
Concentration: ≥25 ng/μl
DNA amount: single library preparation starts from at least 2.5ug, and the total amount should be determined case by case.

Human Whole Genome Resequencing:

Purity: OD260/280=1.8-2.0 without degradation and RNA contamination
Concentration:
≥25ng/μl
DNA amount:
≥1 μg (although more than 2.5 μg is recommended).

Whole Genome Resequencing FFPE Sample Requirements:

Condition: DNA samples without RNA contamination
Purity:
OD260/280=1.8~2.0
Input amount: ≥500N ng; N represents the number of library construction and 2 libraries are recommended.
For sequencing depth 50X, 5 libraries will be needed, but we recommend 6 libraries.

Bisulfite Sequencing:

For the genomic DNA samples you will provide:
Purity: OD 260/280=1.8-2.0, without DNA degradation and protein pollution
DNA amount: single library preparation starts from at least 5μg

MeDIP Sequencing:

For the genomic DNA samples you will be providing:
Purity:OD260/280=1.8-2.0, without degradation and RNA contamination
Concentration: ≥50ng/μl.
DNA amount: single library preparation starts from at least 5μg, the greater the amount the better

ChIP Sequencing:

For the ChIPed DNA samples you will provide:
Purity: OD260/280=1.8-2.0
Concentration: ≥1ng/μl.
Basic Requirements:

  1. Gel electrophoresis analysis result should be provided to make sure that the size of DNA fragments is between 100bp to 500bp, and most of the fragments should be about 250bp.
  2. DNA total amount: ≥10ng (the greater the amount the better) for every single library preparation.

Reduced Representation Bisulfite Sequencing (RRBS):

For the genomic DNA samples you will be providing:
Purity:OD260/280=1.8-2.0, without RNA contamination
Concentration: ≥50ng/μl
DNA amount: single library preparation starts from at least 3μg, the greater the amount the better.
RRBS is available for both human and mouse samples

Metagenomic Sequencing:

Metagenomic sequencing should be discussed case by case. A proposal can be provided after we have enough discussion and communication about your project.

Whole Genome Metagenomic Sequencing:

For the microbial genomic DNA you provide us:
Purity:OD260/280=1.8-2.0
Concentration: ≥50ng/μl
DNA amount: single library preparation starts from at least 5μg, and the total amount should be determined case by case.

16S rDNA Tagging

For the PCR products samples you provides us:
Purity: OD260/280:1.8-2.0
Concentration: ≥50ng/μl
DNA amount: single library preparation starts from at least 5μg, and the total amount should be determined case by case.

Exome Sequencing:

For the genomic DNA samples you will provide us:
Purity: OD260/280=1.8~2.0, without degradation and RNA contamination
Sample concentration: ≥37.5ng/μl
DNA amount: 1μg (2.5μg gDNA recommended).

Whole Exome Sequencing FFPE Sample Requirements:

Condition: DNA samples without RNA contamination
Purity:
OD260/280: 1.8~2.0
Input amount: ≥200 ng; concentration ≥2.5 ng/μl; Main band size ≥500bp OR ≥1ug; concentration ≥10 ng/μl; Main band size ≥250bp.

Target Region Sequencing:

For the genomic DNA samples you will provide us:
Purity:
OD260/280=1.8-2.0, without degradation and RNA contamination
Concentration: ≥37.5ng/μl
DNA amount: 1μg (2.5 μg gDNA recommended).

Genotyping:

For the genomic DNA samples you will provide us:
Purity: OD260/280=1.8-2.0
Concentration: ≥50 ng/μl
DNA amount: single library preparation starts from at least 1μg. DNA quantity demanded for multiple libraries (built N times) 0.5X(N+1) μg.

Optical Mapping:

For the genomic DNA samples you will provide us:
Sample condition: Bacteria DNA with size≥150Kb
Sample concentration: 5-10 DNA/ image
We suggest sending bacterial cultural plate. For bacterial cultural plate, two more control plates are needed for re-culture. Label the culture condition and pathogenicity.
For pathogenic bacteria, we suggest sending Bacteria Plug.

Immune Repertoire Sequencing:

Purity: OD260/280=1.8~2.0, without degradation and RNA contamination
Sample concentration: ≥37.5ng/μl
DNA amount: 2.5 μg ≤ amout < 5 μg (for library construction once); amount ≥ 5 μg (for library construction more than twice).

Human MHC-Seq:

Purity: OD260/280 = 1.8~2.0, without degradation or RNA contamination
Sample concentration: ≥37.5ng/μl
DNA amount: single library preparation starts from at least 1 μg (although ≥2.5 μg is recommended).

RNA

RNA Sample Requirements

Please provide analysis results of the RNA sample using one or several of the following methods: Qubit, NanoDrop, AGE and Agilent 2100.
Please purify samples, avoiding contamination by polycarbonate, protein and exonuclease. Please clearly state the nature of the solvent used in the sample, in the shipment Sample Information Form.

RNA Sequencing (Transcriptome):

Sample condition: Integrate total RNA samples that have been treated with DNase. Avoid protein contamination during RNA isolation.

Common Library Construction Strategy

Sample quantity (for library construction once):

  1. Pant and fungi: total RNA ≥ 20μg
  2. Bacteria: total RNA ≥ 5μg
  3. Mammal (human, rat and mouse): total RNA ≥ 5μg
  4. Other species: total RNA ≥ 10μg

Sample concentration:

  1. Plant and fungi: ≥ 250ng/µl
  2. Bacteria: ≥ 65ng/µL
  3. Mammal (human, rat and mouse): ≥ 65ng/uL
  4. Other species: concentration ≥ 150ng/µl

Sample purity: OD260/280 = 1.8-2.2, OD260/230 ≥ 2.0

  1. Plant and fungi: RNA 28S:18S ≥ 1.0, RIN ≥ 6.5
  2. Bacteria: RNA 23S:16S ≥ 1.0, RIN ≥ 7.0
  3. Animal: RNA 28S:18S ≥ 1.0, RIN ≥ 7.0

Truseq Low-input Library Construction Strategy

Sample quantity (for library construction once):

  1. Pant and fungi: total RNA ≥ 2μg
  2. Mammal (human, rat and mouse): total RNA ≥ 400 ng
  3. Insects: total RNA ≥ 2μg
  4. Other species: total RNA ≥ 2μg

Sample concentration:

  1. Plant and fungi: ≥ 20ng/µl
  2. Mammal (human, rat and mouse): ≥ 5ng/uL
  3. Insects: ≥ 20ng/µL
  4. Other species: concentration ≥ 20ng/µl

Sample purity:

  1. Plant and fungi: RNA 28S:18S ≥ 1.0, RIN ≥ 6.5, OD260/280 ≥ 1.8, OD260/230 ≥ 1.8
  2. Mammal (human, rat and mouse): RNA 23S:16S ≥ 1.0, RIN ≥ 7.0
  3. Other species: RNA 28S:18S ≥ 1.0, RIN ≥ 7.0

RNA Sequencing (Quantification):

Sample condition: Integrated total RNA samples that have been treated with DNase; Avoid protein contamination during RNA isolation
Sample quantity (for library construction): Total RNA ≥ 5 µg
Sample concentration: ≥ 200 ng/µL
Sample purity: OD260/280 = 1.8-2.2; OD260/230 ≥ 1.8; for animal RIN ≥ 7.0, for plant and fungi RIN ≥ 6.5; 28S:18S ≥ 1.0

Quantification FFPE Sample Requirements:

Total RNA: ≥200ng; ≥100ng under the condition of RNA samples after DNAase treatment
RNA concentration:
≥20ng/μl
Purity: OD260/280=1.8~2.2
The FFPE sample can be stored and delivered under room temperature.

small RNA Sequencing:

Sample condition: Integrated total RNA samples. Avoid protein contamination during RNA isolation.

Sample quantity (for every single library construction):

  1. General requirements: total RNA ≥10 μg (standard), total RNA ≥ 200ng (minimum)
  2. For plasma/serum or samples used in Co-IP: total RNA ≥100 ng
  3. For miRNA(< 200nt): RNA ≥1μg

Sample concentration:

  1. General requirements: concentration ≥200ng/μL
  2. For plasma/serum samples: concentration ≥ 2ng/μL
  3. For miRNA(< 200nt): concentration ≥20 ng/μL

Sample purity (for eukaryotes except insects): 28S:18S ≥1.5, RIN ≥8.0

small RNA FFPE Sample Requirements:

Condition: RNA samples after DNAase treatment
Total RNA:
≥2μg; ≥1μg under the condition of RNA samples after DNAase treatment
RNA concentration: ≥20ng/μl
Purity: OD260/280=1.8~2.2

Whole Transcriptome Sequencing:

Sample condition: Integrate total RNA samples (no mRNA isolation). Avoid protein contamination during RNA isolation.
Sample quantity (for library construction once):
total RNA ≥ 5 μg
Sample concentration:
≥ 300 ng/μL
Sample purity (for eukaryotes except insects): 28S:18S ≥1.0, RIN ≥7.0

Immune Repertoire Sequencing:

Purity: RIN ≥ 7.0, 28S/18S ≥ 1.0, the baseline is smooth and 5S peak is normal
Sample concentration: ≥ 200ng/μL
RNA amount: 5 μg ≤ amount <10 μg (for library construction once); amount ≥ 10 μg (for library construction more than twice).

Proteomics

Proteomics Sample Requirements

Sample condition: animal tissue, plant tissue, bacteria, cell, body fluid, impurity-rich samples or samples with low content protein and extracts from protein.

Proteome Profiling:

Sample quantity demanded (single):
Plant tissue:
≥1g wet weight
Animal tissue:
≥100mg wet weight
Bacteria: ≥200mg wet weight
Fungi: ≥2g wet weight
Body fluid:
≥5 ml
Cell (number): ≥5×106 cells
Protein extracts:
≥500 μg (Concentration ≥1 mg/ml)

Quantitative Proteomics:

Sample quantity demanded (single):
Plant tissue: ≥1g wet weight
Animal tissue:
≥100mg wet weight
Bacteria: ≥200mg wet weight
Fungi: ≥2g wet weight
Body fluid:
≥5 ml
Cell (number): ≥5×106 cells
Protein extracts:
≥500 μg (Concentration ≥1 mg/ml)

Phosphoproteomic Analysis:

Sample quantity demanded (single):
Plant tissue: ≥1g wet weight
Animal tissue:
≥100mg wet weight
Bacteria: ≥200mg wet weight
Fungi: ≥20g wet weight
Body fluid:
≥5 ml
Cell (number):
≥5×106 cell number
Protein extracts:
≥500 µg (Concentration ≥1 mg/ml)

Protein Identification:

  • Gel band: the amount of proteins ≥30µg, concentration ≥1µg/µl. The gel band needs to be visible with coomassie blue or silver staining.
  • Gel spot: the amount of proteins ≥50pmol. Salt content: volatile inorganic salt < 20mM, stable inorganic salt < 5mM.
  • Protein molecular weight detection:
    • Sample type: purified protein solution or powder (the purity of the sample should be > 90%)
    • Total amount of protein: ≥50ug
    • Protein concentration: ≥1ug/ul
  • If silver staining method is used, please be sure not to fix the gel with glutaraldehyde.

Target Proteomics:

Sample quantity demanded (single):
Plant tissue: ≥1g wet weight
Animal tissue:
≥100mg wet weight
Bacteria: ≥200mg wet weight
Fungi: ≥2g wet weight
Body fluid:
≥5 ml
Cell (number): ≥5×106 cells
Protein extracts: ≥500 μg (Concentration ≥1 mg/ml)

FFPE

Formalin fixation, paraffin embedding is a conventional storing method for diseased tissue in histopathology. Because formalin-fixed, paraffin-embedded (FFPE) tissues may be stored indefinitely at room temperature and nucleic acids (both DNA and RNA) may be recovered from them decades from the original fixation, FFPE tissues are an important resource for historical studies in medicine.
In the years since our founding, we have greatly expanded our capabilities and now offer you the ability to sequence your FFPE preserved samples. This normally very challenging endeavor enables the study of a wide range of biological and clinical samples and research applications.

To learn more, download the FFPE brochure.

FFPE Sample Requirements (for Human, Mouse and Rat):

 

DNA Level (Whole Exome Sequencing, Target Region Sequencing, Whole Genome Resequencing):

 
WES (Whole Exome Sequencing) and TRS (Target Region Sequencing) FFPE Sample Requirements:

Condition: DNA samples without RNA contamination
Purity:
OD260/280: 1.8~2.0
Input amount: ≥200 ng; concentration ≥2.5 ng/μl
More than 100 folds depth data is recommended.

Whole Genome Resequencing FFPE Sample Requirements:

Condition: DNA samples without RNA contamination
Purity:
OD260/280=1.8~2.0
Input amount: ≥500N ng; N represents the number of library construction and 2 libraries are recommended;Concentration: ≥10 ng/ul; Main band size ≥250bp.
For sequencing depth 50X, 5 libraries will be needed, but we recommend 6 libraries.

 

RNA Level (Quantification and small RNA):

 
For Quantification:

Total RNA: ≥200ng; ≥100ng under the condition of RNA samples after DNAase treatment
RNA concentration:
≥20ng/μl
Purity: OD260/280=1.8~2.2

For small RNA Sequencing:

Condition: RNA samples after DNAase treatment
Total RNA:
≥2μg; ≥1μg under the condition of RNA samples after DNAase treatment
RNA concentration: ≥20ng/μl
Purity: OD260/280=1.8~2.2

The FFPE sample can be stored and delivered under room temperature.