| DNA/RNA Purifi cation
Thermo Fisher Scientifi c
Ratastie 2
01620 Vantaa
Finland
Phone: +358 9 329100
Fax: +358 9 3291 0415
info.microplateinstruments@thermofi sher.com
www.thermo.com/kingfi sher
Simple, Automated Nucleic Acid Purifi cation with the Thermo Scientifi c
KingFisher 96 Magnetic Particle Processor
When processing large numbers of samples, you need a method
that’s fast, easy, and reliable. That’s exactly what you’ll fi nd when
you combine Invitrogen ChargeSwitch
®
DNA and RNA purifi cation
kits with the Thermo Scientifi c KingFisher
®
96 instrument. The
KingFisher
®
96 provides a fast, fl exible and easy-to-use solution
for automated nucleic acid purifi cation in 96-well plate format using
ChargeSwitch
®
kits. Rapid and accurate DNA or RNA purifi cation is
achieved using the innovative approach of shifting magnetic particles
through purifi cation phases instead of liquid. The instrument is
Figure 1—Highly reproducible process. Total RNA from 48 mammalian
capable of processing 96 samples per run, with a volume range
293F cell samples were processed on the KingFisher
®
96 using the
of 20–1,000 µl. For high-throughput needs, the KingFisher
®
96 ChargeSwitch
®
Total RNA Cell Kit. The RNA yield was quantified on a
can be interfaced with liquid handling, robotics, and plate-stacking
NanoDrop
®
spectrophotometer (NanoDrop Technologies). Results indicate
instruments, providing a fully automated solution.
consistently high yields of total RNA (CV = 5.5%, average yield 21.4 ± 1.2).
ChargeSwitch
®
technology provides simple, rapid, and effi cient
isolation of genomic DNA, plasmid DNA, and total RNA from various
sample types. It is a novel magnetic bead-based technology that uses
a pH-dependent ionic switch for the purifi cation of nucleic acids. In
low-pH conditions (pH <6.5), the ChargeSwitch
®
magnetic beads
have a positive charge that binds the negatively charged nucleic
acid backbone. Proteins and other contaminants do not bind and
are washed away in an aqueous buffer. To elute nucleic acids, the
charge on the surface of the beads is neutralized by raising the pH to Figure 2—High-quality genomic DNA from various plant samples. gDNA
8.5 using a low-salt buffer. The purifi ed nucleic acid elutes instantly
was isolated from various plant samples using the ChargeSwitch
®
gDNA
into this buffer and is ready for use in downstream applications.
Plant Kit and the KingFisher
®
96. Purified gDNA was analyzed on agarose
gel. Lane 1-8: pine; corn seeds; corn leaves; sunflower seeds; soybean
leaves; Arabidopsis; tomato seeds; spinach. Lane M: DNA Ladder.
The ChargeSwitch
®
method is ideal for automation on the KingFisher
®

96. High yields of pure DNA and RNA are easily obtained in just 10–40
minutes, depending on the protocol. Optimized, validated purifi cation
protocols can be downloaded from www.thermo.com/kingfi sher or
www.invitrogen.com/naprep.
Reproducible and cross-contamination–free nucleic
acid isolation
Using the ChargeSwitch
®
with the KingFisher
®
96 improves the
speed and accuracy of nucleic acid isolation without compromising
Figure 3—Cross-contamination–free process. The 96-well plate was arrayed
reproducibility. The results clearly demonstrate the performance,
in a checkerboard pattern with 48 human buccal cell and water samples
reproducibility and absence of any cross-contamination during the
(blanks). Genomic DNA was isolated using the ChargeSwitch
®
gDNA Buccal
purifi cation process (Figures 1, 2 and 3 ). Cell Kit with the KingFisher
®
96. Purified gDNA was subjected to PCR and
analyzed on agarose gel. Lane M: DNA ladder; lane (+): buccal cell sample;
lane (–): blank.
Protocol Guide ı 2008 View entire protocol online at www.biotechniques.com/protocol ı BioTechniques ı 19
PGNov08-Thermo-Invitrogen.indd 19 10/25/07 6:53:38 PM
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