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MasterPure Gram Positive DNA Purification Kit

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Contents: Gram-Positive Cell Lysis Solution, MPC Protein Precipitation Reagent, Ready-Lyse Lysozyme, Proteinase K, TE Buffer, RNase A.

MasterPure Gram Positive DNA Purification Kit

Purify genomic DNA from challenging Gram-positive bacteria for a wide variety of molecular biology applications.

Key features

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  • 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

Number of purifications: 100

Item ID MGP04100
TBD
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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.

References

  1. Hoffman, L. and Jarvis, B. (2003) Epicentre Forum 10(3), 3.
  2. Jarvis, B. and Hoffman, L. (2004) Epicentre Forum 11(3), 7.
  3. Luna, V.A. et al. (2006) J. Clin. Microbiol. 44, 2367.
  4. Li, J. et al. (2007) Immunol. 75, 1811.

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