Carrier-iRNA is polyinosinic acid of Molecular Biology Grade.
|C078||Carrier-iRNA, 10 mg/ml||0,5 ml||€ 71,40|
|C079||Carrier-iRNA, 10 mg/ml||5x 0,5 ml||€ 251,58|
For precipitation of small amounts of RNA or DNA (previously called RNA carrier R057)
Carrier-iRNA is polyinosinic acid of Molecular Biology Grade. It is an efficient inert carrier for ethanol precipitation of picograms and higher quantities of RNA or DNA. Carrier-iRNA offers several advantages over other carriers, such as tRNA, yeast RNA, or sonicated DNA, for recovering nucleic acids prior to downstream applications. Carrier-iRNA is synthetic polymer, which is not source of biological contamination in the samples. Nucleic acids recovered after precipitation in the presence of Carrier-iRNA are immediately suitable for downstream applications such as PCR and RT-PCR.
Components and packaging
- Carrier-iRNA is supplied in deionized, ultrapure, and sterile water (18 Mohm.cm) at a concentration of ~10 mg/ml.
- Basic packaging contains 0,5 ml of Carrier-iRNA in 2 ml plastic vials with screw cap.
- Carrier-iRNA is a part of the set for RNA or DNA precipitations, containing besides Carrier-iRNA also Carrier-ACRYL and Carrier-GLY. Comparison of various carriers for RNA or DNA precipitation and key references are shown in Table 1.
Storage and Stability
Store at temperature -20°C ± 5°C. Carrier-iRNA is stable until the expiration date printed on the tube label. To reduce the viscosity after freezing, we recommend heating the Carrier-iRNA tube to 37°C for 15 min.
Each batch of Carrier-iRNA is analyzed in several assays. For the assays, DNA or RNA is examined in the Carrier Assay Buffer (CAB): 10 mM Tris-HCl, 2 mM MgCl2, 1 mM dithiothreitol, pH 7.5 at 37°C.
- Nucleic acid precipitation assay. Economy DNA marker (Cat. No. D071; 2.5 µl) is mixed with 0.2 ml 10 mM Tris buffer, pH 8.0 + 1 mM EDTA, 1 µl Carrier-iRNA, 20 µl of 3 M sodium acetate, pH 5.2, and 0.6 ml of 96% Ethanol. After 30 minutes at 2 - 8°C the mixture is centrifuged for 10 min at 12,000 x g, analyzed by electrophoresis in agarose gel with ethidium bromide and observed under UV light. More than 90% of all components of the DNA marker is recovered in the precipitate.
- Nick activity assay. Plasmid pUC19 (1 µg) in 0.2 ml CAB is incubated with Carrier-iRNA (50 µg) for 1 hours at 37°C, followed by electrophoresis in agarose gel with ethidium bromide. No nicking activity is observed.
- Ribonuclease assay. RNA (1 µg) in 50 µl CAB with Carrier-iRNA (50 µg) is incubated for 1 hours at 37°C, followed by electrophoresis in agarose gel with ethidium bromide. No changes in properties of RNA are observed under UV light.
|Table 1 - Comparison of various carriers for RNA/DNA precipitation.|
Chemically defined RNA, which is more suitable as carrier for cDNA synthesis and other RNA/DNA manipulations than widely used rRNAs or tRNAs.
|Could inhibit reactions catalyzed by terminal transferase or polynucleotide kinase. Interferes with determination of RNA or DNA concentrations.|
Inert neutral carrier, which does not inhibit DNA cloning, DNA-protein interactions, and enzyme reactions. Does not interfere with determination of RNA/DNA concentrations. Does not co-precipitate short oligonucleotides (≤ 20 pbs).
Does not co-precipitate short oligonucleotides (≤ 20 pbs).
Purified glycogen does not inhibit DNA cloning and most enzyme reactions; does not interfere with determination of RNA/DNA concentrations. It is suitable as inert carrier for precipitation of shorter oligonucleotides (≥ 8 pbs).
|May inhibit some DNA-protein interactions and reverse transcription of long RNA templates.|