Description
Purify genomic DNA from challenging Gram-positive bacteria for a wide variety of molecular biology applications.
Contents: Gram-Positive Cell Lysis Solution, MPC Protein Precipitation Reagent, Ready-Lyse Lysozyme, Proteinase K, TE Buffer, RNase A.
Key features
- Purify genomic DNA from challenging Gram positive bacteria for a wide variety of molecular biology applications
- Produces clean, high molecular weight genomic DNA ready for PCR and other molecular biology applications such as restriction digestion, cloning, and Southern blotting
- Scalable method for large sample volumes with no wasting of reagents with smaller samples
- Complete kit includes lysozyme; no separate purchase required
- Robust kit works on a variety of tested species
- Flexible kit also purifies gDNA from Gram-negative bacteria
Product information
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 non-mammalian, non-avian 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 Figure. 1. The DNA purified using the kit is suitable for PCR analysis (Figure. 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 |
| 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.
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| 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 analysed 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. |
SDS
MasterPure Gram Positive DNA Purification Kit
MANUALS
masterpure gram positive dna purification kit