For decades, when flash freezing was not an option the preferred method for fresh tissue collection and protection has been the use of products like RNAlater® to protect the RNA at room temperature. Now there is a better alternative. GenTegra RNAdvantage provides significantly better protection for the RNA in tissues using RIN values as a measure of RNA quality.
RNAdvantage consistently yields RNA with RIN scores that are ≥ 1 better than RNAlater. More importantly the RIN values when using RNAdvantage are at or above the desired RIN value of ≥7, ensuring excellent RNAseq results. Why risk having your RNA extraction rejected for RNAseq because the RIN number is too low? By using RNAdvantage, your RIN values will meet or exceed the recommended RIN number for excellent RNAseq data.
Using GenTegra RNAdvantage is identical to how you use RNAlater. RNAdvantage can be substituted for RNAlater with no changes to your current protocol. The only difference will be that your RIN values for the RNA will be on average 1 RIN number greater. Your RNA will be better protected and ensures your RNA is suitable for RNAseq at all sequencing services, commercial or university core labs.
Benefits
- RIN values ≥ 7 for most tissues
- Rapid RNase inactivation
- Best protection at 7 and 14 days at RT
- No rtPCR or library prep inhibition
- Ensures reliable RNAseq data
- Makes field collection of tissue samples easier
Figure 2.. Agilent TapeStation 4200 gel images of purified mouse tissue RNA samples after 7 and 14-day incubation at room temperature in RNAdvantage or RNAlater solution.
Virtually guaranteed excellent RNAseq results
University core sequencing facilities often have more stringent requirements on the RIN number for RNA being submitted for RNAseq. The RIN values for GenTegra RNAdvantage protected tissue samples even at 14-days of room temperature storage meet or exceed the requirements for commercial sequencing services such as Azenta (Genewiz0 RIN ≥6.0 and Novogene RIN ≥5.8. The RIN values for the RNAdvantage protected samples are either at or so close to the Core lab requirements as to ensure excellent RNAseq results.
Figure 3. In every tissue tested the RIN score at 7-days for RNAdvantage protected tissue exceeded the stringent requirement for a RIN score of ≥7 and in all tissues the 14-day RIN scores were such that excellent RNAseq could be performed on the samples.
Of equal importance with the RIN numbers is that there is no interference with downstream qPCR, etc. As can be seen in Figure 4, RNAdvantage and RNAlater show no significant impact on the expected Ct values when compared to untreated tissue samples.
Figure 4. Average Ct values for GAPDH-qPCR from purified mouse tissue RNA samples after 7-day incubation in RNAdvantage or RNAlater
28s and 18s Ratios The ratio of the 18s peak to the 28s peak is another metric for RNA integrity. Over an assortment of six different tissue types, the ratios for 28s/18s were consistently better for RNA tissue samples protected with RNAdvantage at 14 days then for RNAlater protected samples at 7 days (data available upon request). As an example, spleen tissue showed essentially no change between 7 days and 14 days at room temperature.
Figure 5. Agilent TapeStation 4200 electropherograms (EPGs) for six purified mouse RNA tissue samples after 7 and 14-day incubation at room temperature in RNAdvantage or RNAlater solution.
RNA Protection from Beginning to End™ GenTegra offers a suite of products for RNA protection:
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RNAdvantage for protecting RNA in fresh tissue samples
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RNAssure™ for purified RNA, providing protection as the RNA is eluted
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GenTegraRNA™ for safely shipping or storing RNA at room temperature
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