For the rapid purification of genomic DNA from various bacterial species found in milk
- Genomic DNA can be isolated from as few as 10 bacterial cells in 1 mL of milk
- Isolate genomic DNA from both Gram negative and Gram positive bacteria in milk
- Fast and efficient spin-column format
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Bio-Synthesis's Milk Bacteria DNA Isolation Kit provides a rapid method for the isolation and purification of genomic DNA from the various bacterial species found within milk. The kit allows for the isolation of genomic DNA from both Gram negative and Gram positive bacteria in milk samples.
Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the bacteria present in the milk sample are pelleted and lysed, and the released bacterial genomic DNA is bound to Bio-Synthesis's column in the presence of Binding Solution (BIND). Under these conditions only the genomic DNA will bind to Bio-Synthesis's resin while most of the contaminating RNA and cellular proteinaceous components are removed in the flowthrough. The bound DNA is then washed to remove any remaining impurities (WASH). Lastly, the purified bacterial DNA is eluted into 100 - 200 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.
The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with PCR, quantitative PCR and Southern Blot analysis. |
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FEATURES AND BENEFITS
- Isolate genomic DNA from all types of bacteria found in milk - Bacterial genomic DNA can be isolated from both Gram negative and Gram positive bacteria using the kit.
- Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 45 minutes.
- Isolate high quality DNA - No degradation of the genomic DNA isolated with the Milk Bacterial Genomic DNA Isolation Kit is observed.
- Isolate genomic DNA from milk samples with very low bacterial densities - Genomic DNA can be isolated and detected from milk samples with bacterial concentrations as low as 10 bacterial cells in 1 mL of milk.
- Recovered genomic DNA is suitable for downstream applications - Purified genomic DNA can be used in a number of downstream applications including restriction enzyme digestions, PCR amplifications, quantitative PCR and Southern Blot analysis.
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APPLICATIONS
- Restriction Enzyme Digestions
- PCR
- Quantitative PCR
- Southern Blot Analysis
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| Figure 1. Isolation and Detection of DNA from as Little as 10 Bacterial Cells in 1 mL of Milk. Increasing amounts of E coli (101 , 103 , 105 , 107 ) were added to 1 mL pasteurized milk samples, and the bacterial DNA was subsequently isolated using Bio-Synthesis's Milk Bacteria DNA Isolation Kit with a 200 µL elution volume. One microliter of the isolated genomic DNA was then detected in a 20 µL real-time PCR reaction using primers specific to E coli. Bacterial genomic DNA could be isolated and detected from all the 1 mL milk samples, including the sample that contained only 10 bacterial cells (blue line). |
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Figure 2. End-Point PCR of Milk Bacterial DNA samples. One milliliter of milk was spiked with various amount of E coli. Bacterial DNA was isolated in duplicate using Bio-Synthesis's Milk Bacterial DNA Isolation Kit, and 1 µL of each 200 µeL lution was used for conventional PCR. For analysis 10 µL of each PCR was loaded on an agarose gel. The amount of bacteria in each milk sample is as follows: A = 107, B = 105, C = 103, D = 101. Lane X is a negative control (no template) and Lane M is Bio-Synthesis's PCRSizer 100bp DNA Ladder.
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| Maximum Milk Input |
1 mL |
| Time to Complete 10 Purifications |
45 minutes |
| Minimum Detection Limit |
10 bacteria in 1 mL |
| Bacteria Species Processed |
Gram positive and Gram negative |
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| Milk Bacteria DNA Isolation Kit Contents |
| 1. |
Digestion Buffer |
| 2. |
Lysis Solution |
| 3. |
Binding Solution |
| 4. |
Wash Solution I |
| 5. |
Wash Solution II |
| 6. |
Elution Buffer |
| 7. |
Proteinase K (lyophilized) |
| 8. |
Lysozyme (powder) |
| 9. |
Micro Spin Columns |
| 10. |
Collection Tubes |
| 11. |
Elution Tubes |
| 12. |
Product Insert |
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Long-Term Storage: All solutions should be kept tightly sealed and stored at room temperature. The Lysozyme should be stored at -20°C upon arrival, and the Digestion Buffer should be stored at -20°C after addition of the lysozyme. The lyophilized Proteinase K should be stored at -20°C upon arrival and after reconstitution. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage: For laboratory research use only, not for diagnostic use! Not for use in humans.