Protocols and Applications Guide (US Letter Size) - Promega

||||||||| 9DNA Purification
Promega Publications
PN090 MagneSil® Genomic, Large Volume
System, for large-sample genomic DNA
isolation
(www.promega.com
/pnotes/90/12727_22/12727_22.html)
PN085 Automated 96-well purification of
genomic DNA from whole blood
(www.promega.com
/pnotes/85/10904_07/10904_07.html)
PN083 Expanding the capabilities of plant
genomic DNA purification
(www.promega.com
/pnotes/83/10492_25/10492_25.html)
Citations
Bailey, A.M. et al. (2003) Robotic nucleic acid isolation using
a magnetic bead resin and an automated liquid handler for
biological agent simulants. J. Assoc. Lab. Automation 8,
113–20.
This study describes the development of a system that can
rapidly and accurately detect traces of biological agents
from environmental samples. Using Erwinia herbicola and
Bacillus subtilis var. niger as models for potential biological
warfare agents, a method for DNA extraction using the
Wizard® Magnetic DNA Purification System for Food,
MagneSil® Blood Genomic, Max Yield System, and a
combination of the two was automated on a Beckman
Coulter Biomek® FX robotic liquid handling system. The
isolated DNA was used in a TaqMan® real-time PCR assay
that specifically amplified and identified DNA species. The
ability of the MagneSil®-based DNA purification technology
to eliminate PCR inhibitors was also evaluated. Various
soil samples, surface swabs and air samples were mixed
with bacterial cultures to see if any contaminants present
in the samples inhibited PCR. The authors found that the
modified MagneSil® method described here eliminated
many PCR inhibitors.
E. Maxwell® 16 System
As laboratories try to improve productivity, the need has
increased for easy-to-use, low- to moderate-throughput
automation of purification processes. The Maxwell® 16
Instrument is designed for efficient, automated purification
from a wide range of sample types. The instrument is
supplied with preprogrammed automated purification
methods and is designed for use with prefilled reagent
cartridges, maximizing simplicity and convenience. The
instrument can process up to 16 samples in approximately
30–40 minutes (depending on sample type and method).
Purified concentrated products are high quality and
obtained at high yield to be used directly in a variety of
downstream applications.
Protocols & Applications Guide
www.promega.com
rev. 3/09
The Maxwell® 16 System purifies samples using
paramagnetic particles (PMPs), which provide a mobile
solid phase that optimizes capture, washing and elution of
the target material. The Maxwell® 16 Instrument is a
magnetic-particle-handling instrument that efficiently
preprocesses liquid and solid samples, transports the PMPs
through purification reagents in the prefilled cartridges,
and mixes efficiently during processing. The efficient
magnetic particle-based methodology used by the
Maxwell® 16 Instrument avoids common problems
associated with automated purification systems, such as
clogged tips or partial reagent transfers, which can result
in suboptimal purification processing. Several
Maxwell® 16 reagent kits are available and allow optimal
purification from a variety of sample types including blood,
FTA® paper, formalin-fixed, paraffin-embedded tissue,
bacteria, plant and animal tissue (see Figures 9.15 and 9.16).
Figure 9.15. A panel of murine tissues purified using the
Maxwell® 16 Tissue DNA Purification Kit on the Maxwell® 16
Instrument. Five microliters of genomic DNA was purified from
50mg of the following mouse tissues: Lane 1, brain; lane 2, heart;
lane 3, intestine; lane 4, liver; lane 5, pancreas; lane 6, spleen;
lane 7, 1cm mouse tail clipping; lane 8, 0.5cm mouse tail clipping;
lane L, Lambda DNA/HindIII Marker (Cat.# G1711) All tissue
samples were added directly to the reagent cartridge without
preprocessing.
Figure 9.16. Consistent purification using the Maxwell® 16
Instrument. Five microliters of genomic DNA was purified from
400µl of human whole blood, 1cm mouse tail, 25mg tomato leaf
or 400µl of an overnight culture of Gram-negative bacteria.
Lane L, Lambda DNA/HindIII Marker (Cat.# G1711).
PROTOCOLS & APPLICATIONS GUIDE 9-22