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One
goal of genomic studies is to reveal new biological targets
for drug development by identifying the DNA sequence, which
then enables analysis of the genes, the resulting RNA molecules,
and/or the proteins. To begin, this task requires the isolation
and manipulation of high quality DNA from the sample. Once
isolated, the DNA must be made into a 'library' containing
individual segments that, together, represent each and every
nucleotide in that sample genome. Libraries are often made
by inserting fragments of sample genomic DNA into larger pieces
of DNA from bacteria or yeast that are highly amenable to
laboratory manipulation. The manipulated DNA carrying the
sample insert can then be grown in a controlled system to
increase the yield. For successful genomic sequencing, the
subsequent isolation and purification of the DNA constructs
is critical and can be time-consuming.
Adsorbents
that provide fast and efficient DNA purification are the key
to making this procedure amenable to automation. The exact
nature of the process varies from one manufacturer to the
next, but the basic process is uniform: once the cells are
lysed, DNA is either adsorbed onto a chemically modified silica
matrix contained in 96-well flow-through plates or magnetically
separated using magnetic or transient paramagnetic bead technology.
RNA, proteins and other cellular components are filtered out
initially or washed free in a subsequent step. The multiple
samples of purified DNA are then simultaneously eluted in
a purified form that is ready for batch sequencing.
The adsorbents are available in prepackaged 96-well microplate
kits from a variety of manufacturers, e.g., Qiagen GmbH (Hilden,
Germany), Eppendorf-5 Prime Inc. (Boulder, Colorado), Promega,
Bio-Rad (Hercules, California), Macherey-Nagel (Duren, Germany),
Whatman (Clifton, New Jersey), BD Biosciences Clontech (Palo
Alto, California), Millipore (Bedford, Massachusetts), Sigma-Aldrich
(St. Louis, Missouri), XTRANA Inc. (Broomfield, Colorado)
and many others.
Learn more about the automation
choices available to the drug discovery scientist to perform
nucleic acid purification in a high throughput manner.
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