Chelex-100 and DNA
nospam.net at hgmp.mrc.ac.uk
Sat Feb 15 10:21:41 EST 2003
I am attempting to develop a quick method to extract Trypanosoma (a
blood parasite) PCR-ready DNA from buffy coats. The first try with a
commercial blood extraction kit (using silica filter spin columns) and
subsequent PCR was unsuccessful.
The literature reports various techniques using an incubation in 1-5%
Chelex-100, followed by various steps of heating and centrifugation.
Not being familiar with Chelex-100, I presume that it is used to
chelate the iron and other metals in erythrocytes and the BC, to
remove PCR inhibitors and reduce degradation to DNA. Judging from the
product information, Chelex-100 are bead-shaped resins that are
ionically charged and bind metal ions, leaving DNA in the supernatant.
The SN can then be further processed for use in PCR.
For anyone who is familiar with the chemistry and/or use of
Chelex-100, is this a plausible procedure?
Thanks for any input.
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