MasterPure™ Gram Positive DNA Purification Kit

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Last modified: 2008-07-15 14:57:15

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™ and the Gram Positive Cell Lysis Solution. Ready-Lyse™ Lysozyme is a stable solution of a non-mammalian, non-avian recombinant lysozyme, with high specific activity and no net charge at neutral pH. Thus, there is no waiting to dissolve the lysozyme and it does not bind DNA.

Figure 1 B. subtilis DNA

Figure 2 PCR of BacF/BacR

Figure 1. Electrophoretic Analysis of DNA Purified Using the MasterPure™ Gram Positive DNA Purification Kit. A. DNA from Bacillus subtilis (ATCC 6051) was separated on a 1% agarose gel and stained with SYBR® Gold.Lane 1, kilobase ladder. Lane 2, 300 ng of B. subtilis DNA. B. Pulsed field gel electrophoresis of Bacillus subtilis (ATCC 6051) DNA. Lane 1, Phage lambda ladder. Lane 2, 300 ng of DNA. Lane 3, 600ng of DNA. Figure 2. PCR Product from DNA Purified Using the MasterPure™ Gram Positive DNA Purification Kit. B. subtilis DNA (1ul containing 0.5 ng) was amplified by PCR using forward primer 5'-AGG GTC ATT GGA AAC TGG G and reverse primer 5'-CGT GTT GTA GCC CAG GTC ATA. The cycle conditions were 95°C for 2 min and then 30 cycles of [95°C for 45 sec; 55°C for 45 sec; 72°C for 45 sec] followed by 72°C for 2 min. Lane 1, 100 bp ladder. Lane 2, PCR product from 0.5 ng of B. subtilis DNA. Lane 3, no template PCR.

Last modified: 2008-05-14 16:46:46

Applications

• Obtain genomic DNA from gram positive bacteria for fosmid library construction, PCR, restriction digests or Southern blotting.

Benefits

• PCR-ready DNA
• Scalable method
• High molecular weight DNA
• No extra enzyme to purchase
• Tested on Bacillus, Streptococcus and Listeria

Last modified: 2008-05-14 16:46:46

Protocols for: MasterPure™ Gram Positive DNA Purification Kit

(catalogue number MGP04020 / MGP04100)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Hoffman, L.M. and B.W. Jarvis, (2003) EPICENTRE Forum. 10 (3), 3.
2. Jarvis, B.W. and L.M. Hoffman, (2004) EPICENTRE Forum. 11 (3), 7.

Last modified: 2008-05-14 16:46:46