Recovering DNA from agarose gels with pumice.
نویسندگان
چکیده
منابع مشابه
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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We recently described a method for recovering nucleic acids separated from polyacrylamide gels, which is based on the sensitive on-gel DNA detection with zinc-imidazole followed by passive elution from 32-μm average-size gel microparticles into the elution buffer. The rationale for the preferential use of DNA micropurified by this method, as compared to DNA obtained by a conventional method—bas...
متن کاملSection Ii Resolution and Recovery of Large Dna Fragments
This section describes the application of agarose gel electrophoresis to both analytical and preparative separation of DNA fragments. Standard agarose gels separate DNA fragments from ∼0.5 to 25 kb, whereas pulsed-field agarose gels resolve molecules from ∼10 to >2000 kb. Descriptions of standard and pulsed-field agarose gel electrophoresis as well as parameters affecting resolution of large DN...
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A new method for isolating DNA from agarose gels is described. The method involves the simultaneous transfer of all DNA-fragments from an agarose slab gel onto DEAE-cellulose paper and the elution of the individual fragments from the paper with 1 M NaCl. DNA isolated from agarose gels in this way is susceptible to cleavage with several restriction endonucleases, and can be labeled in vitro with...
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A problem that arises when cloning DNA is the isolation of specific DNA molecules following restriction enzyme digestion or PCR amplification. Typically, a mixture of DNA fragments is electrophoretically separated by agarose gel electrophoresis, the gel is stained with ethidium bromide, and the DNA is visualized by UV transillumination. The DNA fragments of interest are excised from the gel usi...
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ورودعنوان ژورنال:
- BioTechniques
دوره 20 3 شماره
صفحات -
تاریخ انتشار 1996