About Oligo Clean & Concentrator™
The Oligo Clean & Concentrator™ provides a streamlined method for efficient recovery and clean-up of DNA/RNA fragments and oligonucletides ≥16 nt from labeling (radioactive, biotin, DIG, etc.) and other enzymatic reactions. Unincorporated nucleotides, short oligos, dyes, enzymes, and salts are effectively removed by the clean-up procedure.
There is no need for organic denaturants or chloroform. Instead, the kit features Fast Spin column technology and employs a single-buffer system that allows for efficient DNA/RNA adsorption to the matrix of Zymo-Spin™ IC Column. DNA/RNA is washed and concentrated into a small volume of water (≥6 µl). Purified DNA/RNA, available in just 2 minutes, is suitable for hybridization, gel shift assays, enzymatic reactions, ligation, sequencing, microarray analysis, etc.
|Sample Sources||Enzymatic reaction mixtures containing DNA/RNA fragments (radioactive-, biotin-, DIG-labeled) and oligonucleotides ≥16 nt.|
|Product Detergent Tolerance||≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤0.1% SDS.|
|Recovery Volume||Binding capacity of the Zymo-Spin™ IC column is 10 µg of ssDNA/RNA or 5 µg of dsDNA with a typical recovery >90 %. The column can be eluted with ≥6 µl.|
|Compatibility||Single-straded (ss) and double-stranded (ds) DNA and RNA.|
|Purity||High-quality DNA/RNA (A260/A280 nm <1.8; A260/A230 nm <1.8) eluted with water is especially well suited for hybridization, sequencing, ligation, and PCR.|
|Size Limit||For oligonucletides ≥16 nt, up to 23 kb.|
Add 100 µl Oligo Binding Buffer to 50 µl sample1.
Add 400 µl ethanol2 (95-100%), mix briefly by pipetting and transfer the mixture to a provided Zymo-Spin™ Column3 in a Collection Tube.
Note: If required, scale up volumes proportionally.
Centrifuge at ≥10,000 x g for 30 seconds. Discard the flow-through.
For radioactive samples: Transfer the column into a new collection tube then discard the tube containing the radioactive flow-through appropriately.
Add 750 µl DNA Wash Buffer to the column. Centrifuge at ≥10,000 x g for 30 seconds and discard the flow-through. Then centrifuge at maximum speed for 1 minute.
Transfer the column to a microcentrifuge tube. Add 15 µl water4 directly to the column matrix and centrifuge at ≥10,000 x g for 30 seconds to elute the oligonucleotide. Ultra-pure oligonucleotide in water is now ready for use.
The Oligo Clean & Concentrator™ was used to purify small DNA fragments after restriction enzyme digestion, as a part of a CRISPR library generation protocol. A large number of RNA guide libraries were constructed to label repetitive loci in Xenopus egg extracts for an efficient method of whole genome screening.