COMPARATIVE STUDY ON VARIOUS METHODS FOR RECOVERY OF DNA FROM AGAROSE GELS TO EXPLORE MOLECULAR DIVERSITY
Abstract
Objective: Although a few techniques exist for recovery of DNA from agarose gels, they generally require expensive equipment, extended elution times or considerable handling of the sample after elution. The objective of this study is to determine the effective conventional method for maximum recovery of DNA from agarose gels. Methods: The protocols combine different extraction methods such as dialysis tubing, paper strip, spin column and phenol-chloroform were used for a quick and effective recovery with good yield and quality. Results: Using paper strip method, DNA fragments of 800bp from an agarose gel slice were easily recovered. The recovery efficiency was about 70% and the eluted DNA can further be used directly for downstream applications, such as PCR and restriction digestion. Conclusion: This method could also be used to recover large DNA fragments from the agarose gel without degradation since it helps to remove all impurities from agarose gel and it also reduces the clogs during the process.Downloads
References
Yu Z, Morrison M. Improved extraction of PCR-quality community DNA from digesta and fecal samples. Biotechniques 2004;36:808-13.
Oliveira CF, Paim TG, Reiter KC, Rieger A, d'azevedo PA. Evaluation of four different DNA extraction methods in coagulase-negative staphylococci clinical isolates. Revista do Instituto de Medicina Tropical de São Paulo 2014;56:29-33.
Sambrook J, Russell DW. Recovery of DNA from Agarose gels: Electrophoresis onto DEAE-cellulose membranes. Cold Spring Harbor Protocols;2006
Miller DN, Bryant JE, Madsen EL, Ghiorse WC. Evaluation and optimization of DNA extraction and purification procedures for soil and sediment samples. Appl Environ Microbiol 1999;65:4715-24.
Thuring RW, Sanders JP, Borst P. A freeze-squeeze method for recovering long DNA from agarose gels. Anal Biochem 1975;66:213-20.
Wieslander L. A simple method to recover intact high molecular weight RNA and DNA after electrophoretic separation in low gelling temperature agarose gels. Anal Biochem 1979;98:305-9.
Tautz D, Renz M. An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem 198;132:14-9.
Chen CW, Thomas CA. Recovery of DNA segments from agarose gels. Anal Biochem 1980;101:339-41.
Ivanova NV, Dewaard JR, Hebert PD. An inexpensive, automationâ€friendly protocol for recovering highâ€quality DNA. Mol Ecol. Notes 2006;6:998-1002.
Zassenhaus HP, Butow RA, Hannon YP. Rapid electroelution of nucleic acids from agarose and acrylamide gels. Anal Biochem 1982;125:125-30.
Hengen PN. Recovering DNA from agarose gels. Trends Biochem Sci 1994;19:388-9.
Krothapalli U, Nuthi LS, Kumar VP. Biochemical and molecular aspects of DNA in raw and ripen fruit and vegetables. Asian J Exp Sci 2009;23:507-10.
Tabak HF, Flavell, RA. A method for the recovery of DNA from agarose gels. Nucleic Acids Res 1978;5:2321–32.
Chouikh Y, Volovitch M, Yot P. A simple and fast electrophoretic method for elution of nucleic acids from gels. Mol Biol reports. 1979;5:237-9.
Errington J. A rapid and reliable one-step method for isolating DNA fragments from agarose gels. Nucleic acids research. 1990 Sep 11;18(17):5324.
Kumar Vemuri P, Veeravalli S. Expression, Purification and Characterization of Human Recombinant Galectin 3 in Pichia pastoris Iran J Biotechnol 2014;12:3-8.
Chomczynski P, Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on. Nat Protoc 2006;1:581-5.
Published
How to Cite
Issue
Section
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.