Hello,
We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't fully understand what it is used for from my internet search. Is it just a simple DNA purification kit, or is it something more functional? I am interested in recombinant protein production. Can I purify my ligation product with this before cloning into bacteria, or can I purify my PCR product with this before ligation? In which scenarios is this kit indispensable?
Thank you.
https://uk.vwr.com/store/product/4601944/illustratm-microspintm-g-25-columns
These are sephadex spin columns and work on the basis of size exclusion. So dna of more than 10 bases long will flow round the beads and elute early while small salts will have to flow through the pores of the beads and will elute much later. They can be used for dna purification from most smaller molecules and have been used to remove radioactive salts from end labelling of oligos with the labelled oligo eluting first off the column. The G25 is an indication of the size range that can be separated on these columns
Bump Paul Rutland.
Beyond that, as it is a size-exclusion resin, it can be used to separate all sorts of major molecules from their base solutions, hence it's the number one go-to for quick buffer exchange. Thusly, you can also use it for protein buffer exchange in case you may ever need it.
And after you used it, don't just throw it away, just give it a few more bed volumes of buffer and you'll be fine for the next run, as long as you long-term store it in the conventional >=20% ethanol solution so it doesn't spoil.
Illustra™ MicroSpin™ G-25 columns are designed for the rapid purification of DNA, RNA, and other biomolecules from low molecular weight contaminants such as salts, nucleotides, and unincorporated labels. These columns use size exclusion chromatography, where molecules are separated based on their size as they pass through a gel filtration matrix. The G-25 resin inside the columns excludes molecules larger than approximately 5 kDa, allowing them to be eluted quickly, while smaller molecules are retained within the matrix and eluted later.
**Common applications include:**
1. **Desalting:** Removing salts from nucleic acid preparations.
2. **Buffer Exchange:** Changing the buffer composition of a sample.
3. **Purification:** Removing unincorporated nucleotides after labeling reactions.
4. **Rapid Cleanup:** Cleaning up small volumes of biomolecules after enzymatic reactions or synthesis.
These columns are user-friendly and enable quick and efficient sample processing, which is especially useful in molecular biology and biochemical research.
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