RiboShield™ RNase Inhibitor
RiboShield™ RNase Inhibitor는 ribonucleases의 활동을 차단하여 RNA를 RNase digestion 으로부터 안정적으로 보호하는 재조합 단백질입니다.
RT-qPCR, cDNA 합성 및 RNA 시퀀싱과 같은 RNA에 민감한 애플리케이션에 적합하도록 설계되었습니다.
• 뛰어난 보호 기능을 통해 RNA에 민감한 애플리케이션의 성능 향상
• 특히 SARS-CoV-2 검출을 위한 saliva-based tests 에 적합
• eukaryotic RNases 억제 (RNase A, B and C)
• reverse transcriptases, RNA polymerases and Taq DNA polymerase 와 호환
• 65°C에서 최소 30분간 안정
• Ribonuclease and phosphatase free
• 샘플의 장기 보관에 이상적
• cDNA synthesis
• 1-step RT-PCR
• 1-step RT-qPCR
• RNA purification
• RNA sequencing
• In vitro transcription and translation
• Saliva-based diagnostic testing for SARS-CoV-2
Figure 1. Stable at high temperatures
RiboShield™ RNase inhibitor was incubated at the indicated temperatures for 30 minutes. 40U of the inhibitor were then added to 1μg RNA and 5pg RNase A in 5x UltraScript buffer and incubated at 37°C for 30 min. Samples were then loaded on a 1% agarose gel. RiboShield™ RNase Inhibitor can inhibit RNase A at temperatures up to 65°C for at least 30 minutes.
Figure 2. Superior protection against RNase A
RiboShield™ RNase Inhibitor and three competitor products (40U) were incubated with the indicated amounts of RNase A and 1μg RNA in 5x UltraScript buffer at 37°C for 30 min. Samples were then loaded on a 1% agarose gel. L: Ambion RNA Millennium Markers. The RNase inhibitors used were PCR Biosystems’ iboShield™, Promega’s RNasin®, Bioline’s RiboSafe and ThermoFisher’s RNaseOUT™. RiboShield™ RNase Inhibitor offers the greatest RNA protection amongst the inhibitors tested.
How much RiboShield™ should I add to my RNA samples for storage?
For sample storage, we recommend using RiboShield™ RNase inhibitor as a 100x solution (i.e., the concentration of RNase inhibitor in the storage buffer should be 0.4 U/μL). This final concentration can be exceeded without negatively affecting downstream enzymatic reactions but is generally unnecessary unless you suspect RNase contamination.
How to use RiboShield™ RNase inhibitor?
All PCRBIO products for 1-step and 2-step RT-PCR and RT-qPCR, as well as for cDNA synthesis, already contain the RNase inhibitor, so there is no need to add RiboShield™ RNase inhibitor when using these products.
If used with other kits, we recommend adding 1µL (40U) of RiboShield™ RNase inhibitor in a reaction mix of 20µL. In case the source of RNA is known to be rich in RNases, titration may be required to verify the complete protection of the ribonucleic acid.
What can RiboShield™ RNase inhibitor be used for?
RiboShield™ RNase inhibitor is designed for RNA-sensitive applications such as RT-qPCR, cDNA synthesis, and RNA-seq, where even small amounts of RNase can be detrimental to RNA quality and experimental outcome. In addition, it can also be added to extracted RNA to preserve RNA quality in areas with a high likelihood of RNAse contamination or to RNA extraction buffers such as those used in Pereira et al., 2010, to help improve the preservation of RNA integrity as you progress through your workflow.
(Pereira MJB, Behera V, Walter NG (2010) Nondenaturing Purification of Co-Transcriptionally Folded RNA Avoids Common Folding Heterogeneity. PLOS ONE 5(9): e12953. https://doi.org/10.1371/journal.pone.0012953)
What does RiboShield™ RNase inhibitor do?
RiboShield™ RNase inhibitor acts as a shield against the degradation of target RNA which can be caused by ribonuclease contamination of the RNA preparation.
What kind of RNases can be inhibited using RiboShield™ RNase inhibitor?
RiboShield™ RNase inhibitor is a protein-based inhibitor that non-covalently binds and blocks the activity of the most common and troublesome RNases, including eukaryotic RNases of the neutral type (e.g. RNases A, B, and C). It does not inhibit RNases T1, T2, U1, U2, CL3, RNase I and H.
|PB30.23-02||RiboShield™ RNase Inhibitor||2,500 Units||PCRbiosystems||1 x 62.5µL|
|PB30.23-10||RiboShield™ RNase Inhibitor||10,000 Units||PCRbiosystems||4 x 62.5µL|
|제품 자료||RiboShield-RNase-Inhibitor-Flyer-1.pdf||Upload date: 2021.07.12|
|제품 매뉴얼||PB30.23-RiboShield-RNase-Inhibitor-Manual.pdf||Upload date: 2021.07.12|
|제품 MSDS||PB30-Products-MSDS.pdf||Upload date: 2021.07.12|