A simple and cheap one-step method to isolate proteins or nucleic acids by electrophoresis in disposable gel columns is reported. A disposable syringe was modified to host a gel column and an elution chamber. Starting from a crude extract of E. coli, the laboratory-made devise allowed the isolation of the maltose binding protein (MBP) fused to a recombinant allergenic molecule with a molecular mass of 58 kDa, from a mixture of several proteins. Also, plasmid DNA could be isolated from a mixture containing chromosomal DNA and RNA, avoiding the use of organic solvents. Electrophoresis was performed at 150 V, 35°C, pH 8.0 and 8.3 for protein and DNA, respectively. The protein or the DNA obtained showed a yield of 80% and a purity grade of 90%, as estimated by densitometry.

Isolation of proteins and nucleic acids by electrophoresis on disposable gel column

COSTA MA;COLOMBO P;
1996

Abstract

A simple and cheap one-step method to isolate proteins or nucleic acids by electrophoresis in disposable gel columns is reported. A disposable syringe was modified to host a gel column and an elution chamber. Starting from a crude extract of E. coli, the laboratory-made devise allowed the isolation of the maltose binding protein (MBP) fused to a recombinant allergenic molecule with a molecular mass of 58 kDa, from a mixture of several proteins. Also, plasmid DNA could be isolated from a mixture containing chromosomal DNA and RNA, avoiding the use of organic solvents. Electrophoresis was performed at 150 V, 35°C, pH 8.0 and 8.3 for protein and DNA, respectively. The protein or the DNA obtained showed a yield of 80% and a purity grade of 90%, as estimated by densitometry.
1996
Istituto di biomedicina e di immunologia molecolare - IBIM - Sede Palermo
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.14243/9618
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