Electroelution

Electroelutionis a method used to extract anucleic acidor aproteinsample from anelectrophoresis gelby applying a negativecurrentin the plane of the smallest dimension of the gel, drawing themacromoleculeto the surface for extraction and subsequent analysis.^[2] For example,electroblottingandpreparative native PAGEare based upon the sameprinciple.^[3]^[4]

DNA extraction

Using this method,DNAfragments can be recovered from a particular region ofagaroseorpolyacrylamidegels. The gel piece containing the fragment is excised (cut out from the whole gel) and placed in adialysisbag withbuffer.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, usingphenol–chloroform extractionfollowed byethanol precipitation.This method is simple, rapid and yields high recovery (75%) of DNA fragments from gel pieces.^[3]

Protein extraction

Proteins are continuouslyelutedfrom a gel column and separated in different fractions according theirisoelectric point.^[1]Preparative native PAGE yields more than 95% recovery ofmetalloproteins.^[4]

References

^^a ^bKastenholz B, Garfin DE (2010)."Isolation of acidic, basic and neutral metalloproteins by QPNC-PAGE"(PDF).Nature Precedings:1–4.doi:10.1038/npre.2010.4617.1.
^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.
^^a ^bZarzosa-Álvarez, Ana L.; et al. (2010)."Electroeluting DNA Fragments".Journal of Visualized Experiments.43(43): 2136.doi:10.3791/2136.PMC3157863.PMID 20834225.
^^a ^bKastenholz, 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.

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[:1-1] Kastenholz B, Garfin DE (2010)."Isolation of acidic, basic and neutral metalloproteins by QPNC-PAGE"(PDF).Nature Precedings:1–4.doi:10.1038/npre.2010.4617.1.

[2] 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.

[Ana-3] Zarzosa-Álvarez, Ana L.; et al. (2010)."Electroeluting DNA Fragments".Journal of Visualized Experiments.43(43): 2136.doi:10.3791/2136.PMC3157863.PMID 20834225.

[:4-4] 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.

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