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RNeasy Protect Saliva Mini Kit

For stabilization and purification of total RNA from human saliva
  • Immediate stabilization of RNA in saliva
  • Convenient stabilization at room temperature
  • Storage and shipping of saliva without ice or dry ice
  • Archiving of saliva samples at –20ºC or –80ºC
  • Purification of high-quality RNA in minutes

The RNeasy Protect Saliva Mini Kit includes the RNAprotect Saliva Reagent, which stabilizes RNA in saliva samples to preserve the gene expression profile, and the RNeasy Micro Kit, which purifies and concentrates total RNA using specialized RNeasy MinElute spin columns. Stabilized saliva samples can be conveniently stored or shipped at 37°C (1 day), 15–25°C (14 days), or 2–8°C (4 weeks) before RNA purification. RNA is purified from 200 μl saliva and eluted in a volume of just 14 μl.

Cat No./ID: 74324
RNeasy Protect Saliva Mini Kit (50)
$851.00
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RNAprotect Saliva Reagent (50 ml) and RNeasy Micro Kit (50)
The RNeasy Protect Saliva Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Protect Saliva procedure.
1
Minimization of transcript degradation in saliva.

Saliva samples were incubated with or without RNAprotect Saliva Reagent at room temperature for 0 or 7 days. After RNA purification, samples were analyzed using Human Genome U133 Plus 2.0 GeneChip Arrays. Comparison between 0 and 7 days shows a 2-fold decrease in signal intensity for 159 transcripts with unstabilized samples, but for only 38 transcripts with RNAprotect stabilized samples (indicated by red spots). (Data kindly provided by Dr. David Wong, University of California, Los Angeles, CA, USA.)

2
Effective stabilization of beta-actin transcript.
RNAprotect Saliva Reagent stabilized β-actin mRNA levels in saliva for up to 3 days at 25°C. mRNA levels were quantified by real-time RT-PCR using the QuantiTect Probe RT-PCR Kit. Each time point shows the mean CT value from 4 or 6 samples from a saliva pool from 3 donors.
3
Stable mRNA levels of cancer biomarkers.
RNAprotect Saliva Reagent stabilized mRNA levels of oral cancer biomarkers for up to 168 hours. Each time point shows the mean CT value from real-time RT-PCR analysis of 3 samples from a saliva pool from 9 donors. H3F3A: H3 histone, family 3A; OAZ1: ornithine decarboxylase antizyme 1; SAT: spermidine/spermine N1-acetyltransferase; S100P: S100 calcium binding protein P; DUSP1: dual specificity phosphatase 1; IL8: interleukin 8; S100A8: S100 calcium binding protein A8 (an abundant mRNA in saliva; not a cancer biomarker). (Data kindly provided by Dr. David Wong, University of California, Los Angeles, CA, USA.)
Performance
Saliva stabilized with RNAprotect Saliva Reagent can be stored or transported for up to 1 day at 37ºC, up to 14 days at room temperature (15–25ºC), or up to 4 weeks at 2–8ºC, or can be archived at –20ºC or –80ºC (see figure "Effective stabilization of β-actin transcript"). Purified RNA from saliva, as an alternative to blood, allows microarray analysis for research into oral and systemic diseases, and gene expression analysis of RNA biomarkers for cancer by real-time RT-PCR (see figures "Minimization of transcript degradation in saliva" and "Stable mRNA levels of cancer biomarkers").
Principle

The RNeasy Protect Saliva Mini Kit provides a complete RNA protection and isolation solution, from sample harvesting to pure RNA, in one kit. Proven RNeasy technology, combined with the RNA-stabilizing properties of RNAprotect Saliva Reagent, allows purification of high-quality, intact RNA. Saliva contains bacteria in addition to cells and free-circulating nucleic acids of human origin. When saliva is collected from a donor and mixed with RNAprotect Saliva Reagent, the RNA is immediately stabilized and uncontrolled growth of bacteria is prevented, ensuring reliable results in downstream gene expression analysis.  Following stabilization, RNeasy technology simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification. RNA in bacteria is already degrading prior to collection and stabilization in RNAprotect Saliva Reagent, and will be of lower quality than RNA purified from cultured cells and animal tissues.

Procedure

Saliva (200 µl) collected from a donor is immediately mixed with RNAprotect Saliva Reagent to stabilize the RNA in the saliva. Total RNA is purified from stabilized saliva using the RNeasy Micro Kit (see flowchart "RNeasy Protect Saliva procedure"). The saliva sample is first centrifuged, and Buffer RLT is then added to the resulting pellet. After addition of ethanol, the sample is applied to an RNeasy MinElute spin column, where RNA binds to the membrane after centrifugation. Traces of genomic DNA are removed by DNase digestion, and contaminants are washed away in several wash steps. Highly pure RNA is then eluted using RNase-free water in a volume of just 14 µl. RNAprotect Saliva Reagent also stabilizes DNA. Protocols for purification of DNA, or RNA and DNA, from stabilized saliva are available.

 

Applications

Total RNA purified from stabilized saliva using the RNeasy Micro Kit represents a noninvasive alternative to blood analysis. The kit allows research into the identification of RNA biomarkers for:

Oral diseases
Systemic diseases (e.g., cancer)
Features
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 8–14 µl
Format Spin column
Main sample type Saliva samples
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 200 µl
Stabilization Yes
Technology Silica technology
Yield Varies

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Kit Handbooks (2)
For immediate stabilization of total RNA in saliva and subsequent RNA purification
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唾液中のトータルRNAの迅速な安定化およびRNA精製
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