Table of Contents

RNA Amount Requirements

RNA Quality Requirements

RNA Sample Shipment

Options of preparing RNA for shipping

A. RNA in nuclease-free water

B. Freeze-dried RNA

C. RNA in ethanol

Shipping on dry ice

Sample Receipt Confirmation

Disclaimer for low sample amount/quality


RNA sample amount and quality are essential for good data quality and key to the project success. Customer should assess the RNA amount and quality before submitting the samples. Arraystar will always perform sample QC upon sample receipt. The sample QC assessment at Arraystar is final. 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.

 

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.

 

RNA Amount Requirements

 

Required Total RNA Amount per Sample

Microarray

LncRNA microarray

2*-5 ug

Circular RNA microarray

2*-5 ug

LncPathTM Pathway microarray

2*-5 ug

Super-enhancer LncRNA microarray

2*-5 ug

T-UCR microarray

2*-5 ug

miRNA microarray

2*-5 ug

piRNA microarray

2*-5 ug

Agilent Gene Expression (GE) microarray

2*-5 ug

Epitranscriptomic microarray

≥ 5 ug* (for mRNAs or lncRNA)
≥ 10 ug* (for circular RNA)

m6A Single Nucleotide microarray

2*-5 ug

Sequencing

microRNA-seq

2*-5 ug

RNA-seq

2*-5 ug

tRNA-seq

≥ 5* ug

tRF/tiRNA-seq

≥ 5* ug

MeRIP-seq

≥ 100* ug

qPCR Array

nrStar™ Functional LncRNA PCR Array (AS-NR-004)

nrStar™ Canonical Conserved miRNA PCR Array (AS-NR-005)

nrStar™ snoRNA PCR Array (AS-NR-003)

2*-5 ug

nrStar™ tRNA PCR Array (AS-NR-001)

nrStar™ tRF&tiRNA PCR Array (AS-NR-002)

≥ 5* ug

qPCR

 

mRNA-, lncRNA-, circRNA, miRNA-qPCR

2*-5 ug

Serum/plasma circRNA-qPCR

2*-5 ug

serum/plasma miRNA-qPCR

0.2*-5 ug

MeRIP-PCR

≥ 100* ug

MazF-PCR

≥ 5* ug

MeRIP-qPCR (qPCR Validation for Epitranscriptomic Array)

MazF-qPCR (qPCR Validation for m6A Single Nucleotide Array)

5* ug (for lncRNA or mRNA)

10* ug (for circRNA)

LC-MS

tRNA modification LC-MS

10*-15 ug

RNA modification LC-MS for selected/isolated RNA class

≥ 500* ng

*The 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 minimum amount to avoid project delays. If the minimum amount is not obtainable, please consult our technical support by email at support@arraystar.com to make special arrangements.

 

RNA Quality Requirements

 

RNA Quality

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.

Figure 1. (A) RNA integrity by formaldehyde denaturing 1.0% agarose gel electrophoresis. For intact total RNA, the 28S and 18S ribosomal RNA bands should be fairly sharp, intense bands. The intensity of the 28s band should be about twice that of the 18s band. DNA contamination will be evident as a high molecular weight smear or band migrating above the 28s band. For degraded total RNA, the rRNAs will appear smeared, faint, or invisible. Genomic DNA contamination, if any, would show as high molecular weight staining migrating on the top of the gel. (B) RNA integrity by Agilent Bioanalyzer, showing electropherograms, particularly the 18s and 28s rRNA peaks, and RNA Integrity Number (RIN) for total RNAs at varying degrees of degradation. A RIN ≥ 7 is required for best results.

 

RNA Sample Shipment

 

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

 

Options of Preparing RNA for Shipping

A.      RNA in nuclease-free water

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 -80C or in liquid nitrogen. Ship on dry ice.  

B.      Freeze-dried RNA

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.

C.      RNA in ethanol

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.

 

Shipping on Dry Ice

         Use nuclease-free certified, screw-cap, 1.5 mL capacity microtubes;

         Seal the cap with Parafilm strip;

         Place the sample tubes in a plastic bag;

         Use 10 kg dry ice as refrigerant (sufficient for both domestic and international);

         Include a copy of the signed Project Form in a waterproof bag, separate from your samples;

         Check the carrier for dry ice/international shipping requirements;

         If possible, avoid a drop-off date with coming weekends, holidays, such as Thursday or Friday;

         Obtain the tracking number and notify by email the Arraystar representative who is working with you for the project.

 

Sample Receipt Confirmation

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.

 

Disclaimer for low sample amount/quality

 

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.