Name: Oligo Clean-Up and Concentration Kit
Brand: Norgen Biotek Corp.
SKU: 34100
Price: 179.000000 USD
Availability: InStock

Features and Benefits

Cleans and concentrates single-stranded or double-stranded DNA or RNA oligonucleotides larger than 10 bases
Rapid and efficient spin column procedure
No phenol, chloroform or alcohol precipitations are involved
High recovery of up to 90%
Efficient removal of enzymatic reaction buffers and proteins
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid spin column procedure for the purification and concentration of up to 10 µg of oligonucleotides of various sizes. The kit is used for the purification of synthesized oligos, as well as the clean-up of oligos from various upstream enzymatic reactions such as ligation, Poly(A) tailing and end-labeling.

Single or double stranded DNA or RNA oligos larger than 10 bp can be purified with this kit (not recommended for the removal of PCR primers). Oligos of up to 1000 bp have been tested with this kit.

This kit purifies oligos 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. The purified oligos 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.

Details

Supporting Data

Figure 1. 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 right two lanes.

Figure 2. 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.

Figure 3. 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.

Kit Specifications
Column Binding Capacity	10 μg for DNA or RNA
Maximum Column Loading Volume	600 μL
Size of DNA/RNA Purified	> 10 nt, single stranded or double stranded
Maximum Amount of Starting Material	10 μg of DNA or RNA
Minimum Elution Volume	20 μL
Time to Complete 10 Purifications	15 minutes

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.

Component	Cat. 34100 (50 preps)
Buffer RL	30 mL
Wash Solution A	20 mL
Elution Solution A	6 mL
Micro Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Documentation

PROTOCOLS

(34100) Oligo Clean-Up and Concentration Kit - Protocol (50 prep)

SAFETY DATA SHEETS

34100 - Oligo Clean-up and Concentration Kit - SDS

RELATED BLOGS

FAQs

Spin Column

What If a variable speed centrifuge is not available?

If a variable speed centrifuge is unavailable, a fixed speed centrifuge can be used. However, reduced yields may be observed.

What will happen if my centrifugation speed varied from the recommended speed?

Varying the centrifugation speed from the recommended setting could result in a decrease in oligonucleotide yields.

Can I process a different oligonucleotide volume?

Yes, you can. All the solutions included in the sample preparation step of this kit are in a linear relationship to the volume of oligonucleotide sample processed. Make sure that you do not deviate from the ratio specified in the product manual. The solutions are optimized per 50 µL of oligonucleotide sample.

What If I added more or less from the specified reagents’ volume?

Adding less volume may reduce your oligonucleotide yields. Adding more may not affect the oligonucleotide yields EXCEPT if more Elution Solution A was added.
Eluting oligonucleotide in more Elution Solution will result in diluting your yield.

What If I forgot to do a dry spin after my third wash?

If you forget to perform a dry spin after the third wash, your oligonucleotide elution may become contaminated with Wash Solution A. This has the potential to dilute the oligonucleotide yield in your elution and could also interfere with downstream applications.

What If the recovery of the oligonucleotide is poor?

If the recovery of the oligonucleotide is poor, ensure that isopropanol was added during the sample preparation step and ethanol was added to Wash Solution A. Additionally, it is recommended to use the Elution Buffer provided with the kit for optimal recovery.

Why do the purified oligonucleotides not perform well in downstream applications?

The underperformance of your oligonucleotide in downstream applications may be attributed to the following factors:

Insufficient washing of the column.
Ensure that the column is thoroughly washed three times with Wash Solution A during the binding step to eliminate any traces of salt, as residual salt can hinder downstream applications.
Incomplete removal of ethanol.
Make certain that the dry spin under the Column Wash procedure is executed to remove ethanol traces before elution, as ethanol can interfere with various downstream processes.
Use of a different Elution Solution.
If a different Elution Solution was used instead of the provided kit solution, assess it for components that might disrupt the application, such as high salts (including EDTA), detergents, and other denaturants. Verify the compatibility of your chosen elution buffer with the intended downstream use to address any potential interference issues.

Citations

Title	Mapping the ? Integrase bridges in the nucleoprotein Holliday junction internediates of viral integrative and excisive recombination
Citation	PNAS 2014.
Authors	W Tong, D Warren, NE Seah, G Laxmikanthan, GD Van Duyne, A Landy

Title	Nucleoprotein architectures regulating the directionality of viral integration and excision
Citation	PNAS 2014.
Authors	NE Seah, D Warren, W Tong, G Laxmikanthan, GD Van Duyne, A Landy

Oligo Clean-Up and Concentration Kit