Are you sure your RNA never degrades over time?

Cellular RNA provides data-rich biological information about gene expression. Genomic RNA in viruses is used as the basis for viral genetics. Researchers rely on RNA extraction kits to purify and collect their RNA samples for downstream applications including RNA-Seq, qPCR, and other expression profiling techniques.

Keeping RNA samples on ice all the time can slow RNA degradation. However, during handling, the sample temperature can often rise above 0 °C, and even brief exposure to elevated temperature is detrimental to RNA, especially with prevalent contamination from endogenous or environmental RNases1,2. Additionally, the amount of RNase contamination is highly variable depending on sample types and extraction methods. Therefore, it is important to preventively treat the RNA samples with a stabilization agent immediately after extraction to eliminate concerns of RNA degradation for downstream processing and analysis.

Maximum RNA protection is achieved by treating purified RNA samples with GenTegra RNAssure as soon as they are isolated. RNAssure is a robust, reliable protection product that utilizes GenTegra’s proven, patented Active Chemical Protection™ (ACP) chemistry to protect RNA against all RNases as well as exposure to oxidation. RNAssure easily integrates into extraction and purification kits from all major life science manufacturers and can be used directly in downstream applications without the need for further purification.

Benefits

  • Protect RNA integrity during routine sample handling and preparation
  • Protect RNA samples from adverse or accidental exposure to higher temperatures
  • Inactivate all residual RNase activity carried over from purification process or environmental contamination
  • Protect RNA from oxidation damage and non-specific adherence to plastic
  • Integrates seamlessly into standard RNA extraction kit workflows
  • Protected samples can be used directly in downstream applications without the need for further purification

“… during [normal experimental] handling, the sample temperature can often rise above 0 °C, and even brief exposure to elevated temperature is detrimentalto RNA …”

– Nan Su, PhD
President and CSO, Acepix Biosciences

RNAssure easily integrates with your standard RNA purification workflow

RNAssure Elution Tubes replace the manufacturer’s supplied elution collection tubes and integrates seamlessly with standard column-based purification kits and protocols.

Immediate RNA protection workflow
STORAGE CONDITIONLENGTH OF PROTECTION
Room temperature (15-25 °C)3 days
Refrigerator (2-8 °C)2 weeks
Frozen (< -70 °C)> 1 year

RNA is protected from all trace RNases with RNAssure

The amount of RNase contamination varies widely depending on sample types and extraction methods, and it is extremely difficult to assess the level of contamination in an extracted RNA sample. RNAssure increases RNA stability in the liquid state at 37 °C even in the presence of trace amounts of RNase.

RNA integrity is maintained in the liquid state in the presence of increasing amounts of RNase A (left) and RNase I (right) only when protected with GenTegra RNAssure.

HeLa cell RNA (5 μg) was incubated with the indicated amounts of RNase at 37 °C for one hour in the presence (top row) or absence (bottom row) of GenTegra RNAssure.

RNAssure protects RNA from degradation even at very short exposures to RT

Within less than 15 minutes of exposure at RT, unprotected RNA shows noticeable degradation with progressive deterioration of the RNA quality over longer exposure times.

RNAssure protected RNA shows no significant change in RNA quality throughout the 4-hour exposure to RT.

Following short incubations at RT, commercially sourced purified mouse spleen RNA samples were analyzed on a Bioanalyzer™ for both fragmentation length and RIN scores. In the absence of RNAssure (orange), the RNA sample started to show noticeable degradation, after as short as 15 min exposure at RT, and the RNA quality progressively decreased as the exposure time increased. Whereas in the presence of RNAssure (green), the RNA sample was protected with no significant change in RINe value, throughout the 4 h exposure to RT. Unprotected control samples (orange), GenTegra RNAssure protected samples (green).

RNAssure protects RNA from degradation for long-term exposure at RT

In the presence of RNAssure, no significant degradation was observed compared to control RNA for a minimum of 3 days and up to 7 days at RT.

Following 3 or 7 days of incubation at RT, commercially sourced purified mouse spleen RNA samples were analyzed on a Bioanalyzer for both fragmentation length and RINe scores. A. Prior to incubation at RT, control mouse spleen RNA showed no significant degradation. B. After 7 days at 25 °C, RNA showed severe degradation in the absence of RNAssure. C. Whereas, in the presence of RNAssure, no significant degradation was observed compared to control RNA. D. At 25 °C, it is observed that RIN values for the unprotected RNA (orange) showed a dramatic decrease at 3 days, while the RNAssure protected RNA (green) maintained high RINe scores even after 7 days.

1. Precautions for Handling of RNA.
2. The Basics: RNase Control.

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