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Mechanism of Chelex100 cell lysis and DNA extraction

Mechanism of Chelex100 cell lysis and DNA extraction - Microbiology Forum

Mechanism of Chelex100 cell lysis and DNA extraction - Discuss Microbiology Science and Protocols here. Post questions on the study of viruses, fungi, parasites and bacteria here. Microbiology Forum.


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Old 09-25-2006, 02:21 PM
chovek69
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Default Mechanism of Chelex100 cell lysis and DNA extraction



Hi coleagues,

I would be much grateful if someone share some info on how a simple
resin such as Chelex 100 is able to aid in the lysis of Gram+ bacteria
and endospores. For example many authors use chelex-based methods to
extract DNA from Bacillus spores. In the article by A Stein and D
Raoult. A simple method for amplification of DNA from paraffin-embedded
tissues. Nucleic Acids Res. 1992 October 11; 20(19): 5237–5238. is
noted that a simple boiling did not produce PCR amplifiable DNA where
as boiling in Chelex did. I know that the resin binds a large portion
of the PCR inhibitors but how is it supposed to participate in the
lysis step ?

The first two articles that mention chelex 100 as a reagent for DNA
isolation are: de Lamballerie X, Zandotti C, Vignoli C, Bollet C, de
Micco P.A one-step microbial DNA extraction method using "Chelex 100"
suitable for gene amplification. Res Microbiol. 1992 Oct;143(8):785-90.

Walsh, P.S., Metzger, D. A., Higuchi, R., 1991. ChelexТ 100 as a
medium for simple extraction of DNA for PCR based typing from forensic
material. BioTechniques, 4, 506-513.

Unfortunately I have not access to them and can't find em anywhere. If
anybody is able to attach please help.

Ivan

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cell , chelex100 , dna , extraction , lysis , mechanism


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