Carrier-GLY is a highly purified polysaccharide, glycogen, derived from oysters.
|C083||Carrier-GLY||1 ml||€ 50,40|
|C084||Carrier-GLY||5x 1 ml||€ 167,58|
|C085s||Carrier-iRNA, GLY, ACRYL set||1x 0,5 ml + 2x 1 ml||€ 125,58|
For precipitation of small amounts of RNA or DNA
Carrier-GLY is a highly purified polysaccharide, glycogen, derived from oysters. It is an inert carrier, free of host DNA, RNA, nickases, DNases and RNases and is preferred over tRNA, yeast RNA or sonicated DNA as a carrier for the precipitation of nucleic acids, because it is less likely to interfere with downstream applications and is more efficient in RNA and DNA precipitation. Low amounts (picograms/ml) of RNA or DNA oligonucleotides as short as 8 base pairs can be recovered using Carrier-GLY at a final concentration 20 µg/ml.
Components and packaging
- Carrier-GLY is supplied as a highly purified glycogen in deionized sterile ultrapure water (18 Mohm.cm) at a concentration of ~20 mg/ml.
- Basic packaging contains 1 ml of Carrier-GLY in 2 ml plastic vials with screw cap.
- Carrier-GLY can also be obtained as a part of a set for RNA or DNA precipitations, containing besides Carrier-GLY also Carrier-iRNA and Carrier-ACRYL, 1 ml each. Comparison of various carriers for RNA or DNA precipitation and key references are shown in Table 1.
Storage and Stability
At temperature -20°C ± 5°C. Material can be repeatedly defrosted. Unopened vial is stable until the expiration date printed on the label
Each batch of Carrier-GLY 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 (2.5 ul; Cat. No. D071) is mixed with 0.2 ml 10 mM Tris, 1 mM EDTA buffer, 1 ul Carrier/GLY, 20 µl of 3 M sodium acetate, pH 5.2, and 0.6 ml of 96% Ethanol. After 30 min at -20 ± 5°C the mixture is centrifuged 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 200 µl CAB is incubated with Carrier-GLY (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 CAB is incubated with Carrier-GLY (50 µg) in 200 ul CAB 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.
- Absence of nucleic acids. Carrier-GLY (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.|
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.|