Oligo Clean-Up and Concentration Kit
Fast and simple clean-up and concentration of oligonucleotides without the use of phenol
For research use only and NOT intended for in vitro diagnostics.
Oligo Clean-Up and Concentration Kit
Fast and simple clean-up and concentration of oligonucleotides without the use of phenol
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Overview
- 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
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
FAQs
Spin Column
If a variable speed centrifuge is unavailable, a fixed speed centrifuge can be used. However, reduced yields may be observed.
Varying the centrifugation speed from the recommended setting could result in a decrease in oligonucleotide yields.
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.
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.
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.
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.
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 | N1-methyl-pseudouridine is incorporated with higher fidelity than pseudouridine in synthetic RNAs |
Journal | Scientific Reports. 2022 |
Authors | Tien-Hao Chen, Vladimir Potapov, Nan Dai, Jennifer L. Ong & Bijoyita Roy |
Title | A Novel Rolling Circle Amplification-Based Detection of SARS-CoV-2 with Multi-Region Padlock Hybridization |
Journal | Diagnostics. 2022 |
Authors | Rajesh Kumari, Ji Won Lim, Matthew Ryan Sullivan, Rachel Malampy, Connor Baush, Irina Smolina, Howard Robin, Vadim V. Demidov, Giovanni Stefano Ugolini, Jared R. Auclair and Tania Konry |
Title | RNA binding to human METTL3-METTL14 restricts N6-deoxyadenosine methylation of DNA in vitro |
Journal | Biochemistry and Chemical Biology. 2022 |
Authors | Shan Qi, Javier Mota, Siu-Hong Chan, Johanna Villarreal, Nan Dai, Shailee Arya, Robert A Hromas, Manjeet K Rao, Ivan R Corrêa Jr, et al. |
Title | Carbamoyltransferase Enzyme Assay: In vitro Modification of 5-hydroxymethylcytosine (5hmC) to 5-carbamoyloxymethylcytosine (5cmC) |
Journal | bio-protocol. 2022 |
Authors | Weiwei Yang, Nan Dai, Yu-Cheng Lin, William Johnson, Romualdas Vaisvila, Peter Weigele, Yan-Jiun Lee, Ira Schildkraut, Ivan R. Correa Jr, Laurence Ettwiller |
Title | Mapping the λ Integrase bridges in the nucleoprotein Holliday junction intermediates of viral integrative and excisive recombination. |
Journal | Proceedings of the National Academy of Sciences of the USA .. 2014. |
Authors | Wenjun Tong, David Warren, Nicole E. Seah, Gurunathan Laxmikanthan, Gregory D. Van Duyne, and Arthur Landy. |
Title | Nucleoprotein architectures regulating the directionality of viral integration and excision. |
Journal | Proceedings of the National Academy of Sciences of the USA 2014. |
Authors | Seah NE, Warren D, Tong W, Laxmikanthan G, Van Duyne GD, Landy A. |