RNA Collection Kit Oragene®-RNA

Oragene® is a trademark of DNA Genotek Inc, Canada

Application

• mRNA expression profiling
• miRNA research
• Developing "expression signatures" of disease
• Can be used for real time PCR

**Origen of RNA comes from white blood cells and oral epithelial cell
**Before you order, we recommend to use free sample kits to confirm whether you can get your specific RNA by Oragene RNA.

Oragene®-RNA's Benefits

• Sample volume: 2 ml
• Total RNA yield: RNA sufficient for > 320 RT PCR/mL of saliva. (A260/A280: >1.8)
• Sample stability at room temperature: 60 days
• Easy collection, transportation and processing
• Lower collection costs with no phlebotomy required
• mRNA expression levels are stable for months at room temperature

Collection method comparison

	Blood collection	Oragene®-RNA	Other product A	Other product B
RNA stability at room temperature	3 days	60 days	14 days	30 days
Self-collection	-	○	○	○
Easy shipping	-	○	○	○
Low infectious risk	-	○	○	○
Total RNA yield	*>Sufficient RNA for 600RT-PCR/ml blood	#RNA sufficient for>320RT-PCR/ml	Various	-
A260/A280	1.8-2.2	＞1.8	-	-
Pre-purification incubation	90 min	50分	-	-
RT-real time PCR	○	○	○	○

*Assuming 5ng total RNA is used per cDNA reaction prepared, according to the Invitrogen M-MLV Reverse Transcriptase protocol and using 2 ul of cDNA per RT-PCR reaction
#Assuming 6ul of eluted material is used per cDNA reaction prepared according to the Invitrogen M-MLV Reverse Transcriptase protocol and using 2ul of cDNA per RT-PCR reaction.

RNA is ribonucleic acid, plays a role in creating a blueprint for based on the blueprint that is in the DNA protein production.
Oragene®-RNA is able to keep about two months the RNA stability at room temperature.
Because of the possible long-term storage by the room temperature of the specimen because it is a kit of self-collecting can be easily, it is recommended to RNA research.
Oragene®-RNA is easy and safe collecting methods and RNA collection kit long-term storage of the features according to the room temperature not to damage the body.

How to use

RNA can be extracted by the 3rd party’s purification

RNA stability test at room temperature with Oragene®-RNA

This white paper describes the quality, yield, and stability of saliva RNA samples collected using Oragene®-RNA and stored at room temperature up to 8 weeks

Material and method

Saliva samples were collected and purified according to protocols supplied with the Oragene®-RNA kits. In brief, after collection1 the samples were vigorously mixed and 250 μL aliquots were removed at pre‑determined time points ranging from 0 days to 8 weeks. The remainder of each sample was stored at room temperature (~22-24°C). At each time point, the 250 μL aliquot was purified according to protocol2. A fraction of the purified RNA was run on a 1% agarose gel to examine the integrity of ribosomal RNA (rRNA) bands. A small fraction of the sample was used to measure total RNA yield and purity as assessed by absorbance at 260 nm, 280 nm, and 320 nm. Absorbance values were corrected for sample turbidity by subtracting the A320 value from A260 and A280 values. Finally, 6 µL of the purified sample was used to prepare cDNA according the instructions shipped with the Invitrogen M-MLV Reverse Transcriptase (Cat. No. 28025-013) using Invitrogen Random Primers (Cat. No. 48190-011). Real-time PCR assays were performed on a Corbett Rotorgene RG-3000A instrument using 1/100th of the cDNA per 25 µL PCR reaction.

Test result

Collection test

Storage test

Stability test

Real-time PCR signal for each of the 5 specifi c RNA messages did not deviate more than 2 Ct values (Figure 3).
Results summarize the data for 5 gene products of interest: human 18S rRNA, β-actin, β-2-microglobulin, Interleukin-8, and Histatin-3.

Figure 3: Summary of Ct values for 5 genes. RNA from saliva from 8 donors collected and stored up to 8 weeks was purifi ed at given time points. Real-time PCR was used to detect the 5 genes of interest. Ct values at each time point were compared to Ct values obtained from samples purified immediately after collection. The red curve represents the average Ct for all donors.

Conclusion

Oragene®-RNA is a non-invasive RNA self-collection kitthat can be used by untrained subjects to collect high quality RNA.
The RNA collected and purifi ed from saliva using Oragene RNA is high quality, with an average collected A260/A280 of 1.9. Oragene RNA can stabilize total RNA at room temperature for up to 8 weeks as demonstrated by agarose gel electrophoresis.
Specifically, 5 human specific genes were demonstrated by real-time PCR to be stable at room temperature for 8 weeks.

Reference data

• MK-AN-019: Use of Oragene®-RNA Samples in the Development of ReaX™ Mastermix Beads for a Two Step RT PCR
• PD-WP-008: Stability and Yield of RNA collected from saliva using Oragene®-RNA for Expression Analysis

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