MasterPure™ Gram Positive DNA Purification Kit
Applications
- Purification of genomic DNA from Gram-positive bacteria for fosmid library construction, PCR, restriction digests, and Southern blotting.
The MasterPure™ Gram-Positive DNA Purification Kit provides all of the reagents needed to purify DNA from Gram-positive bacteria. These bacteria lyse more readily after treatment with Ready-Lyse™ Lysozyme and the Gram-Positive Cell Lysis Solution included in the kit. Ready-Lyse Lysozyme is a stable solution of a recombinant lysozyme from a nonmammalian, nonavian source. It has high specific activity and does not bind DNA.1,2
Examples of Gram-positive bacteria tested with this kit are listed in Table 1. The size range of DNA purified by the MasterPure Gram-Positive DNA Purification Kit is shown in Fig. 1. The DNA purified using the kit is suitable for PCR analysis (Fig. 2), and other molecular biology applications.
Benefits
References
|
|
Species | Culture Medium | Ready-Lyse™ Incubation | DNA Yield µg/ml |
Bacillus subtilis | Brain-Heart Infusion (BHI) | 30 min | 9.0 |
Listeria monocytogenes | BHI | Overnight | 3.3 |
Staphylococcus aureus | BHI | Not needed | 8.0 |
Streptococcus mutans | Todd-Hewitt | Overnight | 3.0 |
Lactococcus lactis | M17 | 30 min | 1.1 |
Table 1. Incubation times needed for DNA recovery from 1 ml of Gram-positive bacterial culture.
Figure 1. Electrophoretic analysis of DNA purified using the MasterPure™ Gram-Positive DNA Purification Kit. Panel A: DNA purified from B. subtilis (ATCC 6051) was separated on a 1% agarose gel and stained with SYBR® Gold. Lane M, kilobase ladder; lane 1, 300 ng of B. subtilis DNA. Panel B: Pulse-field gel electrophoresis of B. subtilis DNA. Lane M, Phage lambda ladder; lane 1, 300 ng of DNA; lane 2, 600 ng of DNA.
Figure 2. PCR analysis of DNA purified using the MasterPure™ Gram-Positive DNA Purification Kit. B. subtilis DNA was amplified by PCR using BacF/ BacR primers, and the products were analyzed by agarose gel electrophoresis. Lane M, 100-bp ladder; lane 1, PCR product from 0.5 ng of B. subtilis DNA; lane 2, no-template control. |
Ordering
Catalog No. | Size |
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MasterPure™ Gram Positive DNA Purification Kit
MGP04020 | 20 Purifications |
MGP04100 | 100 Purifications |