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.
|Species||Culture Medium||Ready-Lyse™ Incubation||DNA Yield µg/ml|
|Bacillus subtilis||Brain-Heart Infusion (BHI)||30 min||9.0|
|Staphylococcus aureus||BHI||Not needed||8.0|
|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.|