RNA sample amount and quality are essential for good data quality and key to project success. Assess the RNA amount and quality before submitting samples. Arraystar will always perform sample QC upon sample receipt; the sample QC assessment at Arraystar is final.
RNA is very sensitive to RNase contamination and degradation. All RNA work must be carried out in a work area thoroughly decontaminated for RNases. Always use RNase-free labwares and reagents. Wear gloves and lab coat.
Should there be issues about the amount or quality, we will contact you immediately for you to decide whether to replace, proceed, or drop the sample from the study.
Compliance and acceptance requirements for RNA samples
If your RNA samples were extracted from any biohazardous sources, you must disclose before placing your order and shipping your samples. Any infectious agents must be inactivated either during RNA purification procedure or by a specific inactivation method. You must ensure your materials and the shipment free of any contamination risk, including the packaging from inside out. We will evaluate and determine the biosafety compliance and acceptance of the samples.
Recommended total RNA amount per sample for Arraystar tRNA and tRF&tiRNA services
*The recommended minimum amount for each sample is intended for use in the entire experiment for a single attempt, including sample QC. It is recommended to supply twice the recommended minimum amount to avoid project delays.
If the recommended minimum amount is not obtainable, please consult our technical support by email at support@arraystar.com to make special arrangements.
| Category | Service | Recommended Total RNA Amount per Sample |
|---|---|---|
| Sequencing | tRNA-seq | > 2 ug* |
| Sequencing | tRF/tiRNA-seq | > 1 ug* |
| Sequencing | tRNA Charging Seq | > 5 ug* |
| Sequencing | tRNA Modification Seq — m1A/m3C/m1G/m2,2G; TRAC Seq — m7G; HAC Seq — m3C | > 5 ug* |
| qPCR Array | nrStar™ tRNA PCR Array (AS-NR-001) | > 5 ug* |
| qPCR Array | nrStar™ tRF&tiRNA PCR Array (AS-NR-002) | > 5 ug* |
| LC-MS | tRNA/mRNA modification LC-MS | 10*-15 ug |
Recommended RNA quality standards for best results
| RNA Quality | Requirement |
|---|---|
| Purification method (Standard) | By standard TRIzol/RNA precipitation method. By most RNA isolation kits. |
| Purification method (including small RNAs) | By standard TRIzol/RNA precipitation method. By small RNA purification kit. For project containing small RNA classes (sizes < 200 nt) such as miRNA, piRNA, tRNA, tRF/tiRNA, or circRNA, make sure the kit is specified to include the small RNAs (e.g. Qiagen miRNeasy). |
| DNase treatment | Optional to decontaminate gDNA. Required if the sample is also used for qPCR. |
| Solution | Nuclease-free, molecular biology grade water. |
| RNA concentration | > 20 ng/uL by Nanodrop. A lower value may be inaccurate for Nanodrop measurement. |
| Purity by OD260/280 | ~2.0 (acceptable between 1.7 and 2.1) by Nanodrop, a low value may indicate protein contamination. |
| Purity by OD260/230 | > 1.8 by Nanodrop, a low value may indicate organics contamination. |
| RNA integrity | By gel electrophoresis: visible, sharp 18S and 28S rRNA bands (Fig. 1A). By Agilent Bioanalyzer: RIN > 7.0 (Fig. 1B). RNAs well known to be degraded/fragmented and in low amount, e.g. serum/plasma/exosomal/FFPE RNAs, are not checked for RNA integrity. Please see Disclaimer. |
Prepare and ship your RNA samples
Please complete, sign, and email us a Project Form prior to sending your samples. The sample package shall be shipped to the address:
ATTN: Samples Receiving (Project#______ )
Arraystar Inc.
9430 Key West Avenue #128
Rockville, MD 20850 USA
Tel: 888-416-6343
No extra ingredients. Best for compatibility and most purposes. 1) Dissolve the RNA in nuclease-free water. The concentration must be > 20 ng/uL. 2) Store at -80°C or in liquid nitrogen. Ship on dry ice.
Less risk for prolonged international shipment due to dry ice loss. 1) Lyophilize the RNA samples with a freeze drier according to the manufacturer’s instructions. 2) Ship the sample on dry ice or at room temperature.
For added assurance to prevent RNA degradation. May incur higher sample loss during recovery. May contain co-precipitation carrier such as glycogen. 1) Redissolve the RNA in nuclease-free water or TE buffer. The final volume should be 100~300 uL. 2) Add 1/10th volume of 3M NaOAc (pH 5.2); add 2.5 volumes of 100% ethanol. Mix well by vortexing. 3) Store at -80°C or in liquid nitrogen and ship on dry ice.
All shipments and inquiries should reference the Project Number assigned to your project by Arraystar. You will receive notification from us upon receipt of your samples. We’ll notify you if there are any discrepancies between your Sample Submission Form and the contents we receive, or if there are any issues with the physical condition of the samples.
For the best results, the RNA samples must meet the amount and quality requirements. In case of low sample amount or quality, we will contact you for you to decide whether to replace, proceed, or drop the sample from the study. With your consent and approval, it is possible for us to proceed with the experiment. In such a case, you understand the data quality and the success rate may decrease progressively with the lowered sample amount or quality. Although usable data may still be obtainable, we cannot predict for certainty nor do we guarantee the quality and success rate of the data outcomes.
Common questions about RNA amount, quality, and shipment
Contact our technical support team for sample amount and quality questions, or request a quote for your tRNA and tRF&tiRNA project.