Preparative and analytical purification of DNA from agarose.

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Preparative and analytical purification of DNA from agarose.

Two procedures were developed for removing DNA from agarose after electrophoretic separation of DNA fragments according to size. Both involve dissolving the DNA-containing agarose in NaI. The preparative technique uses binding of DNA to glass in the presence of NaI. The method is rapid and convenient, and DNA of all molecular weight ranges can be recovered in high yield and without degradation....

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Isolation and purification of large DNA restriction fragments from agarose gels.

This unit describes methods for recovering and purifying DNA restriction fragments from agarose gels. The first basic protocol describes electroelution of the fragment of interest from standard agarose gels using buffer-filled dialysis bags, followed by concentration and purification using an Elutip column. This approach can be used effectively for fragments of all sizes from 50 to 20,000 bp. E...

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A quick and effective in-house method of DNA purification from agarose gel, suitable for sequencing

Sequencing of DNA fragments (e.g., ITS, 16S, 18S, particular genes, and molecular markers) is increasingly required in studies on microbial diversity, microbial genetic population and phylogeny, sequencing of alleles, and searching for SNPs, among others. The cost of obtaining these DNAs, in quantity and quality for sequencing, is high as it involves special kits to recover DNA from gel after P...

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Protection of DNA during preparative agarose gel electrophoresis against damage induced by ultraviolet light.

Preparative gel electrophoresis of double-stranded DNA usually includes staining the gel with ethidium bromide followed by illumination with ultraviolet (UV-B) light. In this report, DNA isolated from agarose gels was found to be a poor substrate for in vitro transcription, transformation of E. coli and PCR. Inhibition was not caused by enzyme-inhibiting impurities in the agarose gel, but was i...

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A Quick, Cost-Free Method of Purification of DNA Fragments from Agarose Gel

In this short communication we report a quick, cost-free method of purification of DNA fragments from agarose gel. Unlike those procedures that involve commercial kits, this method uses glass wool or absorbent cotton to filter agarose gel during a quick spinning-down of DNA, thus significantly simplifying the routine practice of many molecular biologists and decreasing the cost.

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 1979

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.76.2.615