Sample Preparation |
Rapid Extraction of PCR-Ready DNA and RNA
Order
| Protocol
Applications
- Preparation of DNA from Gram-positive and Gram-negative bacteria.
- DNA can be used for PCR, restriction digests, pulsed-field gel electrophoresis
(PFGE), and optical mapping.
The QuickExtract™ Bacterial DNA Extraction Kit provides a simple method
for extracting DNA from Gram-positive and Gram-negative bacteria. To a washed
pellet of the bacterial sample, add QuickExtract Bacterial DNA Extraction Solution
and Ready-Lyse™ Lysozyme Solution. Ready-Lyse Lysozyme is a proven reagent
with over 200 times the specific activity of egg-white lysozyme. Mix and incubate
for 15 minutes at room temperature. For some species, if lysis is not achieved,
the incubation time can be extended. Samples may optionally be heated at 80°C
for 2 minutes to kill any remaining viable bacteria. The DNA is now ready for
PCR or other downstream applications such as restriction digests, PFGE (pulsed-field
gel electrophoresis), and optical mapping. The kit has been tested on a range
of bacteria (Table 1) and has been shown to produce very long DNA strands (Fig.
1).
Benefits
- Single-tube system.
- No toxic organic solvents.
- Suitable for high-throughput applications.
- Ready-Lyse™ Lysozyme supplied.
- Long DNA generated.
Table 1. Species of bacteria that have been tested with the QuickExtract™ Bacterial
DNA Extraction Kit.
| Bacillus subtilis |
E. coli |
| Bifidobacterium spp |
Salmonella typhimurium |
| Brevibacterium linens |
Vibrio gazogenes |
| Clostridium perfringens |
|
| Lactobacillus plantarum |
|
| Listeria monocytogenes |
|
| Staphylococcus equorum |
|
| Streptococcus agalactiae |
|
| Streptococcus pyogenes |
|
| Streptococcus thermophilus |
|
|
|
Figure 1. Long DNA strands from E. coli extracted using the
QuickExtract™ Bacterial DNA Extraction Kit. Many strands
are 400-500 kb. Image courtesy of Dr. Shiguo Zhou, University of Wisconsin-Madison. |
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