// Workers AI · dad joke modeWhy did electroelution go to therapy? It had a shocking separation issue.
Electroelution[1] is a method used to extract a nucleic acid or a protein sample from an electrophoresis gel by applying a negative current in the plane of the smallest dimension of the gel, drawing the macromolecule to the surface for extraction and subsequent analysis.[2] For example, electroblotting is based upon the same principle.[3]
DNA extraction
[edit]Using this method, DNA fragments can be recovered from a particular region of agarose or polyacrylamide gels. The gel piece containing the fragment is excised (cut out from the whole gel) and placed in a dialysis bag with buffer. Electrophoresis causes the DNA to migrate out of the gel into the dialysis bag buffer. The DNA fragments are recovered from this buffer and purified, using phenol–chloroform extraction followed by ethanol precipitation. This method is simple, rapid and yields high recovery (75%) of DNA fragments from gel pieces.[3]
Protein extraction
[edit]Preparative native PAGE yields more than 95% recovery of metalloproteins. Proteins are continuously eluted from a gel column and separated in different fractions according to their isoelectric points.[2]
References
[edit]- ↑ Seelert H, Krause F (2008). "Preparative isolation of protein complexes and other bioparticles by elution from polyacrylamide gels". Electrophoresis. 29 (12): 2617–36. doi:10.1002/elps.200800061. PMID 18494038. S2CID 35874355.
- 1 2 Kastenholz, B (2004). "Preparative Native Continuous Polyacrylamide Gel Electrophoresis (PNC‐PAGE): An Efficient Method for Isolating Cadmium Cofactors in Biological Systems". Analytical Letters. 37 (4). Informa UK Limited: 657–665. doi:10.1081/al-120029742. ISSN 0003-2719. S2CID 97636537.
- 1 2 Zarzosa-Álvarez, Ana L.; et al. (2010). "Electroeluting DNA Fragments". Journal of Visualized Experiments. 43 (43): 2136. doi:10.3791/2136. PMC 3157863. PMID 20834225.
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