Isolation of DNA from agarose gels using DEAE-paper. Application to restriction site mapping of adenovirus type 16 DNA.
نویسندگان
چکیده
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 E coli DNA-polymerase I, T4 DNA-polymerase and T4 polynucleotide kinase. We have used the method to construct restriction endonuclease maps of adenovirus type 16 DNA.
منابع مشابه
Prostate-specific antigen expression in neoplastic human myeloid cell lines.
Sons, 1989:3.2. 5. Chomczynski P, Sacchi H. Single-step method of RNA isolation by acid guanidinum thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162:156–9. 6. Bauer D, Warthoe P, Rohde M, Strauss M. Detection and differential display of expressed genes by DDRT-PCR. PCR Methods Appl 1994;4:S97–108. 7. Bassam B, Caetano-Anolles G, Gresshoff P. Fast and sensitive silver staining of ...
متن کاملCharacterization of avian adenovirus type-4 causing hydro-pericardium syndrome in broilers in Karachi, Pakistan
Avian adenovirus was isolated from naturally infected birds during 1990 to 2003 in the vicinity ofKarachi, Pakistan. The virus is known to cause hydro-pericardium syndrome (HPS) in poultry. The mortalitywas recorded 20-50% in commercial broilers. Fifty field samples were collected; however, six samples wereselected for molecular studies. These isolates were grown and characterized by cell cultu...
متن کاملLimitations of comparative detection of proteins via epitope tagging.
glass plate) was covered by Saran wrap, labeled by hot ink, and exposed 10 min to autoradiography film. The region corresponding to the fragment of interest was cut with a scalpel, the Saran wrap was removed, and the sandwich with the piece of the DEAE paper and Whatman 3MM paper was prepared as for unlabeled DNA fragment. The rest of the procedure was identical to that for the unlabeled DNA fr...
متن کاملModeling of Partial Digest Problem as a Network flows problem
Restriction Site Mapping is one of the interesting tasks in Computational Biology. A DNA strand can be thought of as a string on the letters A, T, C, and G. When a particular restriction enzyme is added to a DNA solution, the DNA is cut at particular restriction sites. The goal of the restriction site mapping is to determine the location of every site for a given enzyme. In partial digest metho...
متن کاملIsolation 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...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleic acids research
دوره 8 2 شماره
صفحات -
تاریخ انتشار 1980