RNeasy Protect Saliva Mini Kit (50)

Protocol tips

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	For stabilization, use approximately 10 µl of saliva Follow manufacturer’s instructions

Protocol tips
For stabilization, use approximately 10 µl of saliva Follow manufacturer’s instructions

Publication protocol

Stabilization of saliva: To stabilize saliva for use with the Qiagen RNeasy Protect Saliva Kit®, 500 µl of RNAprotect Saliva Reagent® was aliquoted into 2 ml Eppendorf tubes. RNAprotect Saliva Reagent® is supplied with the Qiagen RNeasy Protect Saliva Mini Kit® and is designed to halt gene expression changes, limit microbial overgrowth and inhibit destructive RNases. The manufacturer’s directions suggest that 200 µl of saliva be placed directly in 1 ml of the RNAprotect Saliva Reagent®; however, we modified this protocol for use in newborns. Approximately 10 µl of saliva was placed in 500 µl of RNAprotect Saliva Reagent® by flushing the collection syringe with the reagent 4–5 times to ensure that all saliva was removed from the syringe. The Eppendorf tubes then were vortexed, placed immediately on ice and stored at 4° C until total RNA extraction.
Extraction of salivary RNA: Total RNA extraction using the Qiagen RNeasy Protect Saliva Mini Kit® was performed according to the manufacturer’s instructions. The kit uses a silica-membrane RNeasy spin column capable of binding up to 100 μg of total RNA. Each sample underwent treatment with DNase to destroy any residual genomic DNA that would interfere with RNA amplification on the RT-qPCR platform with the use of Qiagen’s RNase-free DNAse Set according to the manufacturer’s instructions. Fresh 70 and 80% ethanol solutions were used during each extraction procedure. The final elution volume of extracted total RNA was approximately 12 µl. Samples were stored at −80° C until further analysis.

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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Protect Saliva Mini Kit (50) below.

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