IBI Tri-Isolate RNA Pure Kit – 100 Reactions
- Sample Size: up to 5 x 106 cultured cells, 10 – 50 mg tissue, up to 200 µl body fluids (blood, buffy coat, serum, plasma), 1 x 109 bacterial cells, 20 – 50 mg plant tissue
- Cost effective phenol, guanidine isothiocyanate solution plus spin column system
- No chloroform phase separation
IBI Tri-Isolate is a phenol and guanidine isothiocyanate plus spin column system for convenient purification of high quality total RNA from a variety of samples including cultured cells, tissue, body fluids, bacteria, and plant tissue.
For more details click on Protocol or Technical Information below.
- No isopropanol RNA precipitation
- No phenol carryover
- High quality RNA: A260/A280 > 1.8, A260/A230 > 1.8
- Operation Time: 15 minutes or less
- Elution Volume: 25 – 50 µl
- Binding Capacity: 50 µg RNA from > 25 µl DNase/RNase-free water
- Applications: cDNA Library Construction, Cloning, RT-PCR (Endpoint), Real-Time PCR, Nuclease Protection Assays, Northern Blotting
Initially, samples are homogenized in IBI Isolate Reagent without chloroform phase separation or isopropanol RNA precipitation. Following sample homogenization, simply bind, wash, and elute the high quality, total RNA in RNase-free water. The purified RNA can be used in a variety of downstream applications, including cDNA library construction, cloning, RT-PCR (End-point), Real-Time PCR, nuclease protection assays, northern blotting.
Tri-Isolate is tested on a lot-to-lot basis. 10 µl from a 50 µl eluate of purified RNA is analyzed by electrophoresis on a 0.8% agarose gel.
Can frozen blood samples be used with the Tri-Isolate kit? Yes. Place the frozen sample into a 1.5ml micro-centrifuge tube and add 3 volumes of Tri-Isolate reagent and allow the sample to thaw. Incubate an additional 5 minutes at room temp and proceed with rest of protocol.
Can the protocol be altered to accommodate high lipid samples such as brain tissue? Yes. After homogenization, centrifuge the sample at 12,000 – 16,000 x g for 2 minutes to remove insoluble particles. After centrifugation of hi-fat sample, a layer of fat will float on the supernatant. Remove and discard the fatty layer than transfer the supernatant to a new 1.5ml centrifuge tube. Continue with RNA binding step #2.