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Sample Requirements
The following information provides general guidelines for the required sample amount when requesting next-generation sequencing (NGS) analysis.
If your sample quantity or quality does not meet the criteria listed below, analysis may still be possible. Please consult with us in advance.
Sample Requirements
| Application | Sample Type | Total Amount | Concentration | Volume | Notes | ||
| Human Genome Sequencing |
Whole Genome Sequencing | PCR-Plus | DNA |
≥ 500 ng | ≥ 10 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 |
| PCR-Free | ≥ 4 µg | ≥ 50 ng/µL | ≥ 30 µL | ||||
| Whole Exome Sequencing | ≥ 2 µg | ≥ 10 ng/µL | ≥ 30 µL | ||||
| Whole Genome Sequencing (Non-Human) | PCR-Plus | DNA | ≥ 500 ng | ≥ 10 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | |
| PCR-Free | ≥ 4 µg | ≥ 50 ng/µL | ≥ 30 µL | ||||
| Microbial Genome Sequencing | Long-Read | High-Molecular-Weight DNA | ≥ 10 µg | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 ・Quality Notes: See *4 |
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| Short-Read | PCR-Plus | DNA | ≥ 500 ng | ≥ 10 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | |
| PCR-Free | ≥ 4 µg | ≥ 50 ng/µL | ≥ 30 µL | ||||
| Small-Scale Sequencing (NGS Petit) | DNA | ≥ 4 µg | ≥ 40 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | ||
| Purified Long-PCR Products | ≥ 4 µg | ≥ 40 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | |||
| Purified PCR Products (up to 800 bp) |
～ 2 µg | ～ 20 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 ・The required total amount may vary depending on the size of the PCR product |
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| GRAS-Di® Genotyping | DNA | ≥ 50 ng | ≥ 2 ng/µL | ≥ 20 µL | |||
| Repeat Motif Detection | DNA | ≥ 4 µg | ≥ 40 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | ||
| Gene Expression Analysis (Reference-Based) | Eukaryotes | Total RNA | ≥ 600 ng | ≥ 20 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | |
| mRNA | ≥ 100 ng | ≥ 3 ng/µL | ≥ 30 µL | ||||
| Prokaryotes | Total RNA | ≥ 1.5 µg | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | ||
| mRNA | ≥ 100 ng | ≥ 3 ng/µL | ≥ 30 µL | ||||
| Isoform Sequencing (full-length mRNA-seq) |
Eukaryotes | Total RNA | ≥ 1.5 µg | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | |
| De novo Transcriptome Sequencing | Eukaryotes | Total RNA | ≥ 600 ng | ≥ 20 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | |
| mRNA | ≥ 100 ng | ≥ 3 ng/µL | ≥ 30 µL | ||||
| Prokaryotes | Total RNA | ≥ 1.5 µg | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | ||
| mRNA | ≥ 100 ng | ≥ 3 ng/µL | ≥ 30 µL | ||||
| Small RNA-Seq | Total RNA | ≥ 2 µg | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | ||
| small RNA | ≥ 60 ng | ≥ 2 ng/µL | ≥ 30 µL | ||||
| Microbial Community Analysis | From DNA Extraction | Sludge | – | – | 5 ～ 10 mL | ・Storage/Transport Temperature: Refrigerated | |
| Soil | 1 ～ 5 g | – | – | ・Storage/Transport Temperature: Refrigerated | |||
| Feces | 0.2 ～ 0.5 g | – | – | ・Storage/Transport Temperature: Frozen | |||
| Saliva | – | – | 0.2 ～1 mL | ・Storage/Transport Temperature: Frozen | |||
| Other | Please inquire | ||||||
| From 1st PCR | DNA | ≥ 150 ng | ≥ 5 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | ||
| From 2nd PCR | Purified 1st PCR Products | ≥ 5 ng | ≥ 0.2 ng/µL | ≥20 µL | ・Concentration Measurement Method: See *2 ,*3 ・Recommended Size: See *5 |
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| From Sequencing | Purified 2nd PCR Products | ≥ 150 ng | ≥ 5 ng/µL | ≥20 µL | ・Concentration Measurement Method: See *2 ,*3 ・Recommended Size: See *5 |
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| Shotgun Metagenomic Sequencing | PCR-Plus | DNA | ≥ 500 ng | ≥ 10 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 | |
| PCR-Free | ≥ 4 µg | ≥ 50 ng/µL | ≥ 30 µL | ||||
| Metatranscriptome Sequencing | Total RNA | ≥ 1.5 ug | ≥ 50 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *1 | ||
| ChIP-Seq | Purified ChIP-DNA | ≥ 20 ng | ≥ 1 ng/µL | ≥ 30 µL | ・Concentration Measurement Method: See *2 ,*3 ・Recommended Size: 100 – 500 bp |
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| CRISPR Screening | Purified 1st PCR Products | ≥ 5 ng | ≥ 0.2 ng/µL | ≥20 µL | ・Concentration Measurement Method: See *2 ,*3 ・Recommended Insert Size : 200 – 400 bp ・gRNA Region: Within 100 bp on the 5′ side |
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| Illumina Amplicon Sequencing | Purified 1st PCR Products | ≥ 5 ng | ≥ 0.2 ng/µL | ≥20 µL | ・Concentration Measurement Method: See *2 ,*3 ・Recommended Size: See *5 |
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| PacBio Amplicon Sequencing |
DNA | Please inquire | |||||
| Purified Long-PCR Products | |||||||
*1 Spectrophotometric measurement is generally acceptable. However, pre-checking with an Agilent 2100 BioAnalyzer or equivalent is recommended.
*2 Spectrophotometric measurements may detect contaminants such as free nucleotides, leading to inaccurate values. We recommend agarose gel electrophoresis for comparison with known DNA standards.
*3 As spectrophotometers may not provide accurate measurements, we recommend commercial dsDNA quantification kits such as the Invitrogen Qubit Fluorometer and Quant-iT dsDNA Kit.
*4 PacBio Sample Requirements:
– DNA should be dissolved in Tris Buffer or Nuclease-Free Water.
– Only double-stranded DNA is accepted; single-stranded DNA cannot be used for library preparation.
– Avoid multiple freeze-thaw cycles.
– OD260/280 ratio should be between 1.8 and 2.0.
– Ensure the sample is free from insoluble materials and RNA contamination.
– The sample should not be exposed to intercalating fluorescent dyes or UV radiation.
– Avoid contamination from tissue components such as heme, humic acid, polyphenols, and polysaccharides.
– Ensure the sample is free from chelating agents (e.g., EDTA), divalent metal cations (e.g., Mg2+), denaturants (e.g., guanidine salts, phenol), and detergents (e.g., SDS, Triton-X100).
*EDTA can be removed during library preparation.
*5 Amplicon Sequencing Requirements:
– For MiSeq 300PE sequencing, we recommend an insert size of approximately 300-450 bp. With our designated linker sequences, the total length of the 1st PCR product will be around 370-520 bp, and the 2nd PCR product (with Illumina adapter sequences) will be around 420-570 bp.
– For NovaSeq/HiSeq 150PE sequencing, we recommend an insert size of approximately 200-250 bp. With our designated linker sequences, the total length of the 1st PCR product will be around 270-320 bp, and the 2nd PCR product (with Illumina adapter sequences) will be around 320-370 bp.
*6 For PacBio amplicon sequencing, the required sample amount depends on the amplicon size. Please contact us for details.
*7 For Iso-Seq, the sample amount listed above applies to comprehensive analysis targeting all mRNA in the sample. If selecting specific mRNA for size selection, different sample amounts may be required. Please contact us for details.
Sample Submission Guidelines
– Samples from human subjects (excluding commonly used cultured cell lines) must be anonymized in compliance with ethical guidelines set by the Ministry of Health, Labour and Welfare of Japan.
– We conduct a quality check on submitted samples before proceeding with analysis.
* If samples does not meet our recommended quality criteria, we may request resubmission or discontinue analysis.
* Twice quality checks per analysis request are included in the analysis fee. Additional checks beyond the second attempt will incur additional charges.