Oligo (Single and Double Stranded RNA and DNA Oligonucleotides) Clean-Up and Concentration Kit

Product Description

• Cleans and concentrates single-stranded or double-stranded DNA or RNA oligonucleotides larger than 10bp
• Rapid procedure – Clean and concentrate 10 oligonucleotides samples in 10 minutes
• No phenol, chloroform or alcohol precipitations are involved
• High recovery of up to 90%
• Efficient removal of enzymatic reaction buffers and proteins
• Complete column purification - The RNA or DNA oligonucleotides are column cleaned and/or concentrated with no need to use the labor-intensive PAGE-based purification.
• Provides high quality oligonucleotides - The purified oligonucleotides are of the highest quality and can be used in a number of downstream applications.

Norgen’s Oligo Clean-Up and Concentration Kit provides a rapid, simple and efficient procedure for the purification and concentration of up to 10 µg of oligonucleotides. The kit is used for the purification of synthesized oligonucleotides, as well as the clean-up of oligonucleotides from various upstream enzymatic reactions such as ligation, Poly(A) tailing and end-labeling. Single or double stranded DNA or RNA oligonucleotides larger than 10 bp can be purified with the kit, therefore the kit is not recommended for the removal of PCR primers. The kit purifies oligonucleotides from other reaction components including proteins, buffers and nucleotides without the use of phenol, chloroform, alcohol precipitation or urea PAGE extraction. The kit provides a high quality product with up to 90% recovery.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the sample containing the oligonucleitide is first mixed with the provided Binding Solution, and the nucleic acid is then bound to Norgen's column (BIND). Under these conditions only the nucleic acid will bind to Norgen's resin while any contaminating proteins or nucleotides are removed in the flowthrough. The bound nucleic acid is then washed to remove any remaining impurities (WASH). Lastly, the purified oligonucleotides are eluted into the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart below.

Workflow

Norgen's Oligo Clean-Up and Concentration Kit provides a high quality product with up to 90% recovery. The purified oligonucleotides can be used in a number of downstream applications including end-point or quantitative reverse transcription PCR, Northern blotting, in situ hybridization, and RNAi studies.

Integrity of Purified DNA Oligonucleotides. Oligonucleotides of different sizes (15mer, 20mer, 42mer and 61mer) were purified using Norgen´s Oligo Clean-Up and Concentration Kit, and the integrity of the oligonucleotides before and after cleaning were compared by running a 15% urea-PAGE gel. Equal volumes of the input (I) and cleaned (C) oligonucleotide were run, and as it can be seen the purified oligonucleotides were of a high quality and integrity. Please note that the kit can also clean a mixture of different oligo sizes, as shown on the far left two lanes.

Integrity of Purified dsDNA. Samples of Norgen`s MiniSizer DNA ladder (25bp-650bp) were purified using Norgen´s Oligo Clean-Up and Concentration Kit, and the integrity of the dsDNA before and after cleaning was compared by running aliquots on a 2% agarose gel. Both the input volume and the elution volume were maintained at 50 µL, and 5 µL and 10 µL of the input and 5 µL and 10 µL of the elution were run on the gel for visual comparison. As it can be seen, the purified dsDNA was of a high quality and integrity. The overall recovery of the different ladder bands is 82% of the input.

Integrity of Purified RNA. Samples of Norgen`s 100b RNA ladder (100b-1000b) were purified using Norgen´s Oligo Clean-Up and Concentration Kit, and the integrity of the RNA before and after cleaning was compared by running aliquots of the input and elution on a 1.4% formaldehyde-agarose gel. Both the input volume and elution were maintained at 50 µL, and 5 µL and 10 µL of the input and 5 µL and 10 µL of the elution were run on the gel for visual comparison. The kit provides efficient RNA recovery of all the ladder bands. The overall recovery of the different ladder bands is 84% of the input.