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Materials for testing of RNA coronavirus, causing
COVID-19
Carrier-ACRYL
For precipitation of small amounts of RNA or DNA
Description
Carrier-ACRYL is linear polyacrylamide (LPA) of Molecular Biology Grade. It is an efficient inert carrier for ethanol precipitation of picograms and higher quantities of RNA or DNA. Carrier-ACRYL offers several advantages over other carriers, such as tRNA, yeast RNA, or sonicated DNA, for recovering nucleic acids prior to downstream applications. Carrier-ACRYL is synthetic polymer, which is not source of biological contamination in the samples. Presence of Carrier-ACRYL during ethanol precipitation results in complete recovery of fragments larger than 20 base pairs while failing to precipitate shorter fragments and free nucleotides. This feature makes Carrier-ACRYL useful for separating reactions products from unincorporated nucleotides and oligonucleotide primers. Nucleic acids recovered after precipitation in the presence of Carrier-ACRYL are immediately suitable for downstream applications such as PCR, RT-PCR, restriction digestion, ligation, and sequencing.
Technical data
Components and packaging
- Carrier-ACRYL is supplied in deionized, ultrapure, and sterile water (18 Mohm.cm) at a concentration of ~25 mg/ml.
- Basic packaging contains 1 ml of Carrier-ACRYL in 2 ml plastic vials with screw cap.
- Carrier-ACRYL is a part of the set for RNA or DNA precipitations, containing besides Carrier-ACRYL also Carrier-iRNA and Carrier-GLY, 1 ml each. Comparison of various carriers for RNA or DNA precipitation and key references are shown in Table 1.
Storage and Stability
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For short terms (months) at temperature 4°C ± 3°C.
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For long terms at temperature -20°C ± 5°C.
Quality control
Each batch of Carrier-ACRYL 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 37oC.
- 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-ACRYL, 20 µl of 3 M sodium acetate, pH 5.2, and 0.6 ml of 96% Ethanol. After 30 minutes at 2 - 8oC 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-ACRYL (50 µg) for 1 hours at 37oC, 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-ACRYL (50 µg) is incubated for 1 hours at 37oC, followed by electrophoresis in agarose gel with ethidium bromide. No changes in properties of RNA are observed under UV light.
- Absence of nucleic acids. Carrier-ACRYL (200 µg) is loaded on agarose gel with ethidium bromide. After electrophoresis, no bands are observed under UV light.
| Table 1 - Comparison of various carriers for RNA/DNA precipitation. | ||
| Carrier | Advantages | Disadvantages |
| Polyinosinic acid |
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. |
|
Linear polyacrylamide |
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). |
|
Polysaccharide (glycogen from oysters)
|
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. |
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Product information
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