Technique / Molecular Biology / DNA analysis techniques / DNA extraction purification and quantification


DNA extraction and DNA purification methods



DNA extraction and DNA purification methods

Generally speaking, strategies for DNA extraction and DNA purification depends on your downstream applications, e.g. southern blot hybridization using high molecular weight genomic DNA, DNA fragments for subcloning, or DNA for PCR etc. Some considerations for DNA extraction and DNA purification:

1. Avoid nuclease contamination (DNase contamination).
2. Avoid shearing of intact DNA molecules.
3. Avoid chemical contamination that can interfere with downstream applications.

Common steps for DNA extraction and DNA purification:

1. Cell or tissue lysis containing detergents (SDS, triton X-100 etc), proteinase K, RNase etc.
2. DNA extraction step (avoid protein, RNA and chemical contamination).
3. DNA precipitation using 75-80% ethanol, isopropanol, PEG or TCA.

Some commonly used DNA extraction and DNA purification methods:

1. Salting out and DNA Precipitation (Guanidinium isothiocyanate).
2. Organic Solvents DNA extraction and DNA precipitation (phenol, chloroform).
3. Silica Resin-Based DNA purification.
4. Anion Exchange-based DNA purification

Last update 31-Dec-2006, Rating of 9 votes.

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By Graciela G. Perez Villaseñor on 03-Sep-2008
I need to kow how DNA purification for preparative DNA probes. DNA from hepatocarcinogenesis tissue.
! will agree if you have some techiques.
Ph. D. Graciela Pérez Villaseñor
Universidad Autonoma Metropolitana. México.
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By M. Rodham on 23-Jul-2008
I wish to know more information about the PUREGENE method of DNA extraction. Any help would be appreciated.
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By aien on 17-Jul-2008
DNA extraction consist of 3 basic steps,which is homogenization,deproteinization and precipitation. can u elaborate the 3 steps?
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By Ijay on 16-Jul-2008
Indeed this is wonderful, please I need protocol for DNA extraction, purification and characterisation. Thanks
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By Bizarreufo on 10-Oct-2007
After adding the solution2 you should not stand for more than 2min or the DNA will damege!!
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By P. kaur on 28-Aug-2007
i am doin dna extraction with phenol chloroform method but shearing of dna is there . how can i aviod that? what are the critical steps, one should take care of?
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By britya on 17-Aug-2007
excellent i want protocol for dna quantificationa and for purifying the same
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By mazrina on 04-May-2007
dear Sir

can u give me DNA extraction and DNA purification methods
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By khawrami on 31-Dec-2006
all related procedure and protocols to RT-PCR used in viral detection.
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