Next-Generation Sequencing Services Order Preparation and Submission
General Sample Requirements
| Sample Type | Volume | Concentration | Amount (Qubit®) | A260/280 Ratio | A260/230 Ratio | RIN |
| gDNA (for WGS) | >30 µl | Illumina >10 ng/µl PacBio >50 ng/µl | Illumina>1 µg PacBio >5 µg | >1.8 | >2 | - |
| gDNA (for amplicon sequencing) | >30 µl | >10 ng/µl | >300 ng | >1.8 | >2 | - |
| Total RNA (for mRNA or lnRNA) | >30 µl | >20 ng/µl | >1 µg | >1.8 | >2 | > 7.0 |
| Total RNA (for *miRNA/ sRNA) | >30 µl | >20 ng/µl | >2 µg | >1.8 | >2 | > 7.5 |
| Total RNA (for Iso-Seq) | >50 µl | >30 ng/µl | >3 µg | >1.8 | >2 | > 7.5 |
| Total RNA (for meta-transcriptome) | >30 µl | >20 ng/µl | >3 µg | >1.8 | >2 | > 7.0 |
*For small RNA analysis, total RNA must be prepared using the appropriate method to ensure that the small RNA fraction is not removed during the purification process. E.g. many column-based procedures have a cut-off size that will eliminate all small RNA, and are not suitable for this purpose. All quantification for the DNA and RNA quantities must use fluorometric methods (e.g. Qubit). If a spectrophotometer (e,g. Nanodrop) is used, we suggest submitting twice the requested amount of samples due to the unreliable detection of this measurement method. Prior to confirmation of your NGS inquiry, you shall receive confirmation of the specific sample requirement according to your ordered library preparation.
DNA/ RNA Samples
| DNA | 1. Lyophilize the DNA to ship on ambient temperature |
| 2. Pack with ice packs/blue ice (2-8 °C) | |
| 3. Use the cold-chain transportation system (2-8 °C) of the courier | |
| 4. DNAstable (Biomatrica) and ship on 2-8 °C | |
| 5. Pack in dry ice (-60 °C to -80 °C) *Recommended. | |
| RNA | 1. Lyophilize the RNA to ship on dry ice |
| 2. Suspend RNA in 75% ethanol and ship on dry ice | |
| 3. RNAstable (Biomatrica) and ship on 2-8 °C | |
| 4. Pack in dry ice (-60 °C to -80 °C) *Recommended. |
It is highly recommended that RNA samples be shipped in dry ice packaging. Other packaging/ transportation methods may add impurities or cause slight degradation of the total RNA. The required quantity of dry ice and ice bags varies with seasons (i.e., weathers), transit time and the thickness of Styrofoam box and receptacle. Please contact your local courier office to estimate the transit time, so that you can estimate the amount of dry ice or blue ice required to ship your order. Normally, dry ice is consumed (sublimates) at a rate of 5 kg per day.
Pre-made Library Pools
Please only submit pre-made libraries after getting confirmation about your order acceptance. Illumina Sequencing Library Concentration: 2 nM-30 nM, quantified by qPCR. Library Size: insert + adapters (120 bp) ± 50 bp (this size estimation does not apply to small RNA library). Library Quality: single main peak, no multiple peaks, no adapter contamination, and no primer dimers.
| Sequencing Strategy | Library Size (insert + adaptors (120 bp) +/- 50 bp for Optimal Results |
| SE 50 | 130 bp~650 bp |
| PE 150 | 320 bp~520 bp |
| PE 250 | 400 bp~650 bp |
| PE 300 | 400 bp~720 bp |
Sample requirement for NGS grade DNA/RNA extraction
| Type | Required | Recommended | Total Biological Replicates |
| Animal tissue | 200mg | 1g | 3 |
| Plant tissue | 2g | 5g | 3 |
| Fungus/Bacterium | Pellet from 10mL overnight fresh culture | Pellet from 30mL overnight fresh culture | 3 |
| Soil/ Stool/ Sludge (gDNA only) | 300mg | 500mg | 3 |
| Other Sample Types | Please download here or contact us to enquire. | ||
Collect sample freshly. Snap freezes each sample with liquid nitrogen. Store and ship with dry ice.