G 50 columns

Manufactured by Roche

The G-50 columns are laboratory equipment used for size-exclusion chromatography. They are designed to separate molecules based on their size and molecular weight, allowing for the purification and isolation of proteins, nucleic acids, and other macromolecules from complex mixtures.

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Lab products found in correlation

Shrimp alkaline phosphatase, by Promega (2 mentions) Adenosine 5 γ 32p triphosphate, by PerkinElmer (2 mentions)

2 protocols using g 50 columns

Plasmid Digestion and Radiolabeling

Cited 1 time

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Plasmids (5 µg) were digested with appropriate restriction enzymes followed by dephosphorylation of the ends with shrimp alkaline phosphatase (SAP) (Promega). The desired fragments were gel purified and radiolabeled with 30 units of polynucleotide kinase (NEB) and 50 µCi of adenosine 5′-[γ-³²P]triphosphate (PerkinElmer). The labeled fragment was purified through G-50 columns (Roche Diagnostics Corporation). Further details were as described previously (17 (link)).

Jha J.K., Li M., Ghirlando R., Miller Jenkins L.M., Wlodawer A, & Chattoraj D. (2017). The DnaK Chaperone Uses Different Mechanisms To Promote and Inhibit Replication of Vibrio cholerae Chromosome 2. mBio, 8(2), e00427-17.

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Radiolabeling and EMSA Assay

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Plasmids (5 μg) were digested with restriction enzymes EcoRV and HpaI, and the resulting DNA ends were de-phosphorylated with shrimp alkaline phosphatase (SAP) (Promega). The desired fragments were gel purified, radio-labeled with 30 units of polynucleotidyl kinase (15 (link)) and 50 μCi of adenosine 5′-[γ³²P] triphosphate (Perkin-Elmer), and purified by passing the mixture through G-50 columns (Roche Diagnostics Corporation). Electrophoretic mobility shift assay (EMSA) was done, as described (15 (link)).

Jha J.K., Ghirlando R, & Chattoraj D.K. (2014). Initiator protein dimerization plays a key role in replication control of Vibrio cholerae chromosome 2. Nucleic Acids Research, 42(16), 10538-10549.

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