Requirements relative to Sequencing projects
To guarantee the normal processing of your project, samples should meet the standards given below. If your samples do not meet these standards, and you are unable to produce higher-quality samples, please contact us before shipping your samples.
- Input quantity should be determined by Qubit® instead of by NanoDrop™
- Samples not meeting these specifications should be designated as “at risk” by the customer, and will be subject to billing regardless of data quality.
|Library type||Sample Type||Recommended amount||Amount required||Volume||Concentration||RNA integrity number (Agilent 2100)||Purity|
|Eukaryotic RNA-Seq||Total RNA (Animal)||≥ 2 μg||≥ 1 μg||≥ 20 μL||≥ 50 ng/μL||≥ 6.8, smooth base line||No degradation, no contamination|
|Total RNA (Plant and Fungus)||≥ 2 μg||≥ 1 μg||≥ 20 μL||≥ 50 ng/μL||≥ 6.3, smooth base line|
|Prokaryotic RNA-Seq||Total RNA||≥ 6 μg||≥ 3 μg||≥ 20 μL||≥ 50 ng/μL||≥ 6.0, smooth base line|
|Eukaryotic Long non-coding RNA Sequencing||Total RNA (Animal)||≥ 4 μg||≥ 2 μg||≥ 20 μL||≥ 50 ng/μL||≥ 6.8, smooth base line|
|Total RNA (Plant and Fungus)||≥ 4 μg||≥ 2 μg||≥ 20 μL||≥ 50 ng/μL||≥ 6.3, smooth base line|
|Eukaryotic small RNA Sequencing||Total RNA (Animal)||≥ 6 μg||≥ 3 μg||≥ 20 μL||≥ 50 ng/μL||≥ 8, smooth base line|
|Total RNA (Plant and Fungus)||≥ 6 μg||≥ 3 μg||≥ 20 μL||≥ 50 ng/μL||≥ 7.5, smooth base line|
|Library type||Sample Type||Recommended amount||Amount required||Volume||Concentration||Purity|
|Human Whole Genome/Exome Sequencing||Genomic DNA||≥ 2 μg||≥ 1 μg||≥ 20 μL||≥ 20 ng/μL||No degradation, no contamination|
|Genomic DNA (PCR-Free)||≥ 3 μg||≥ 1.5 μg||≥ 20 μL||≥ 20 ng/μL|
|Whole Genome Bisulfite Sequencing||Genomic DNA||≥ 6 μg||≥ 3 μg||≥ 20 μL||≥ 50 ng/μL|
|ChIP-Seq||ChIP-Seq DNA||≥ 100 ng||≥ 50 ng||≥ 10 μL||≥ 20 ng/μL||Main peak of 100 bp – 500 bp|
Pre-Quality Control (QC) Instructions
Customers must provide the results of analysis of sample quality obtained using one of the following methods: Qubit®, NanoDrop™, agarose gel electrophoresis, or Agilent 2100. It is recommended samples be analyzed by Qubit/PicoGreen/gel electrophoresis (with quantity indicator), NanoDrop™ quantification is discouraged. If NanoDrop™ is utilized for pre-QC quantification, we strongly recommends that you send more DNA/RNA for processing than the amounts given above.
For gel electrophoresis, the following conditions are recommended:
- DNA: 1.0% agarose gel; 1.0% TAE solution; 100V for 40 min
- RNA: 1.0% agarose gel; 0.5× TBE solution; 180V for 16 min
Note: Different electrophoresis conditions may generate a different, and potentially misleading, QC report on your samples. Therefore, it is highly recommended that you adhere to the conditions recommended above for the initial check, and that you provide a picture of the gel.
Sample Labeling Recommendations
- It is important to prevent the sample labels from being dissolved by solvents and from falling off the tubes. The use of a waterproof marker pen to write directly on the tube wall or the lid is strongly recommended.
- Please make sure that the sample information provided in the project description wizard matches the labels on the tubes.
- Your sample names must consist of up to 8 alphanumeric characters (or underscores) and shall start with a letter. Non-standard names may be amended.
Sample Packing Recommendations
- For DNA and RNA samples, we recommend 1.5 ml or 2 ml screw-cap DNase- and RNase-free microcentrifuge tubes. Please use Parafilm to seal each tube before packaging. We do not recommend shipping samples dissolved in organic solvents (such as absolute ethanol or isopropanol) because the solvents may cause leakage of the samples, which can result in cross-contamination between samples. If it is unavoidable to ship samples in organic solvents, please use screw-cap tubes and seal the opening of the tube with at least 10 layers of Parafilm.
- In order to avoid crushing during shipping, it is highly recommended to place the sample tubes in a container such as a 50ml tube or a box with interior racks/holders. Cotton and absorbent papers can be used to prevent tubes from moving around inside the container.
- RNA samples should be kept in dry ice during shipment. Genomic DNA samples should be kept in blue ice during shipment.
- In order to stick with our high quality control standards, 96-well plates and PCR stripe tubes are NOT acceptable containers for your sample shipping. The only container we allow for sample shipping is 1.5 ml or 2 ml tube.
Disclaimer: The information below only constitutes a recommendation for shipping samples classified as “non-regulated materials”. Due to continuing changes in regulations, customers should always check with their safety office and/or shipping department to ensure regulatory compliance.
- Ensure that all samples conform to our quality standards and that they are prepared and packaged according to the guidelines given above.
- Print the waybill generated when submitting your project and enclose it in the shipment.
- include any QC data for the samples if available (Qubit/Nanodrop/agarose gel electrophoresis/Agilent 2100).
- Use express services (fastest delivery offered), such as DHL, FedEx or UPS. Please make sure that the carrier can facilitate the importation of DNA or RNA samples and dry ice packing (if applicable) to the country of destination.
- Complete the invoice (commercial invoice, customs invoice, or pro-forma invoice) as required for customs.
- “RNA or DNA Samples for Research Use Only”
- Describe the contents as “non-hazardous, non-contagious research sample”
- No Commercial Value, Value for Customs Only
- Declare the value of the goods for customs to be $1.
- Number of samples and volumes
- Type of container
- Sample transportation options:
- Lyophilize the DNA for shipping at ambient temperature
- Pack with ice packs/blue ice (2-8 °C)
- Use the cold-chain transportation system (2-8 °C) of the courier
- DNA Stable (Liquid format, Biomatrica)
- Pack in dry ice (-60 °C – -80 °C)
- Lyophilize the RNA for shipping at 2-8 °C or ambient temperature
- Suspend RNA in 75% ethanol and ship on dry ice
- RNAstable (Biomatrica)
- Pack in dry ice (-60 °C – -80 °C)
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 RNA.
The quantity of dry ice and ice bags needed varies with the season (i.e., room temperature), transit time, and the thickness of Styrofoam box and receptacle. Please contact your local courier office for estimated transit time. Normally, dry ice is consumed (sublimates) at a rate of 5 kg (11 pounds) per day.
Qubit is a trademark of Life Technologies and Thermo Fisher Scientific. NanoDrop is a trademark of NanoDrop Technologies LLC. Agilent 2100 Bioanalyzer is a trademark of Agilent Technologies.