RNeasy Protect Animal Blood System
For collecting and stabilizing animal blood and purifying RNA and miRNA
The RNeasy Protect Animal Blood System provides a complete solution for preparing high-quality RNA from animal blood. Blood from mouse, rat, and other small animals is conveniently collected in RNAprotect Animal Blood Tubes, which contain a reagent that immediately stabilizes cellular RNA. Total RNA is then purified using the RNeasy Protect Animal Blood Kit. Please note that the RNeasy Protect Animal Blood Kit and the RNAprotect Animal Blood Tubes are ordered separately.
Alternatively, the RNeasy Protect Animal Blood Kit can be used to obtain microRNA (miRNA), either in a total RNA fraction or in a separate small RNA fraction. For purification of total RNA containing miRNA, Buffer RWT (cat. no. 1067933) is also required. For purification of small RNA (containing miRNA) and large RNA in 2 separate fractions, the RNeasy MinElute Cleanup Kit (cat. no. 74204) is also required. The procedures can be fully automated on the QIAcube Connect.
Blood samples collected in RNAprotect Animal Blood Tubes can be conveniently shipped at ambient temperature or easily stored in a refrigerator. Samples can be stored at 15–25°C for up to 48 hours, at 2–8°C for up to 2 weeks, or at –20°C for at least 3 months with no significant RNA degradation (see figure "Highly intact RNA"). Collection of animal blood in RNAprotect Animal Blood Tubes prevents changes in transcript levels, ensuring reliable gene expression analysis in applications such as real-time RT-PCR and microarray analysis (see figure "Effective transcript stabilization"). The RNeasy Protect Animal Blood Kit delivers reproducible yields of RNA, and requires less than 30 minutes of hands-on time (see figure "Reproducible RNA yields"). Proven RNeasy technology delivers purified RNA that is highly pure, with virtually no genomic DNA contamination.
When blood samples are collected, the gene expression profile can change significantly within minutes. Anticoagulants such as EDTA prevent blood clotting, but do not prevent changes in gene expression due to induced transcription or regulated turnover of mRNA. Collection of animal blood in RNAprotect Animal Blood Tubes prevents changes in transcript levels. Following digestion and homogenization of the stabilized blood, RNA is then purified from the samples using well-established RNeasy silica-membrane technology (see figure "RNeasy Protect Animal Blood flowchart").
Blood samples are collected in RNAprotect Animal Blood Tubes. The tubes contain a reagent that lyses blood cells and stabilizes intracellular RNA. The RNAprotect Animal Blood Tubes are centrifuged to pellet the samples, which are then washed with water and resuspended in Buffer RSB. After digestion in Buffer RBT with proteinase K, the samples are homogenized by centrifugation through QIAshredder spin columns. Ethanol is added to the samples, and the samples are then centrifuged through RNeasy MinElute spin columns. The bound total DNA is subjected to DNase digestion and washed with Buffer RW1, followed by Buffer RPE. Pure RNA is eluted in Buffer REB. The purification of RNA with the RNeasy Protect Animal Blood Kit can be fully automated on the QIAcube. A separate protocol is provided for isolation of total RNA including miRNA and requires wash Buffer RWT (ordered separately; cat. no. 1067933). For purification of small RNA (containing miRNA) and large RNA in 2 separate fractions, the RNeasy MinElute Cleanup Kit is also required.
Total RNA and miRNA purified with the RNeasy Protect Animal Blood System can be used for many downstream applications, including:
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