+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Isolation and purification of large DNA restriction fragments from agarose gels



Isolation and purification of large DNA restriction fragments from agarose gels



Current Protocols in Immunology Chapter 10: Unit 10.5



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. Electrophoresis directly onto NA-45 paper is also described and provides relatively high yields for fragments smaller than 2000 bp. Protocols are also provided for separating fragments larger than 1000 bp using low gelling/melting agarose gels and purified by phenol extraction, b-agarase digestion of the gel, or via glass beads extraction. Sieving agarose gels can also be used to resolve very small DNA fragments. Removing linkers from a fragment using a column rather than a gel is included, followed by a method for estimating DNA concentrations in solution.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 054013675

Download citation: RISBibTeXText

PMID: 18432696

DOI: 10.1002/0471142735.im1005s08


Related references

An easy and efficient procedure for the isolation of pure DNA restriction fragments from agarose gels. Biochimica et Biophysica Acta 520(3): 498-504, 1978

Simultaneous purification of multiple, large DNA fragments from agarose gels. Biotechniques 17(4): 644, 648-9, 1994

Isolation of large dna fragments from agarose gels using agarase. Trends in Genetics 5(2): 41-42, 1989

An improved method for the purification of large DNA fragments from agarose gels using Wizard Plus SV columns. Analytical Biochemistry 269(1): 218-219, 1999

Dependence of the ligation efficiency of large DNA fragments isolated from agarose gels on the purification method. Preparative Biochemistry and Biotechnology 28(3): 235-241, 1998

Cloning, restriction digestion and DNA labeling of large DNA fragments (greater than or equal to 1 kb) in the presence of remelted SeaPlaque GTG agarose gels. Biotechniques 11(6): 784-6, 788, 790-1, 1991

Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes. Febs Letters 53(1): 84-86, 1975

Purification of DNA fragments from lyophilized agarose gels. Nucleic Acids Research 23(23): 4926-4927, 1995

An Apple II program to estimate sizes of DNA restriction fragments on agarose gels. Journal of Heredity 82(1): 84-85, 1991

Purification of DNA fragments from agarose gels using glass beads. Methods in Molecular Biology 58: 237-240, 1996

Purification and cloning of DNA fragments fractionated on agarose gels. Molecular Biotechnology 3(2): 135-138, 1995

Accurate molecular weight determinations of deoxyribonucleic acid restriction fragments on agarose gels. Biochemistry 22(26): 6180-6185, 1983

A rapid procedure for isolation of dna fragments from agarose gels. Biotechniques: 66-68, 1984

Analysis and purification of synthetic DNA fragments with NuSieve agarose mini-gels. Biotechniques 6(4): 296, 298, 1988

A method for isolation of small DNA fragments from agarose and polyacrylamide gels. Analytical Biochemistry 293(1): 138-139, 2001