For the rapid purification of total DNA from bacteriophages.
For research use only and NOT intended for in vitro diagnostics.
Details
Supporting Data
Figure 1. Effective Host Genomic DNA Removal without Reducing Phage DNA Yield.
Total DNA was isolated from four enriched phage cultures using Norgen's Phage DNA Isolation Kit. A DNase I pre-treatment was performed prior to adding the provided Lysis Buffer. Briefly, 20 units of DNase I was added to 1 mL of enriched phage culture and the mixture was incubated at room temperature for 20 minutes. After the DNAase I treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 4 cultures using Norgen’s Phage DNA Isolation Kit without performing the DNase I treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel. As it can be seen, the phage DNA was safely protected from the DNase I treatment by its coat protein, while the host genomic DNA was efficiently degraded by the DNase I. Thus the DNase I pre-treatment resulted in less host gDNA contamination in the final phage elution without influencing the total phage DNA yield. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)
Figure 2. Optional Proteinase K Treatment Improves DNA Yield for Certain Phage Strains.
Total DNA was isolated with and without the optional Proteinase K treatment using Norgen's Phage DNA Isolation Kit. Briefly, 4 µL of Proteinase K (20 mg/mL) was added to 1 mL of enriched phage culture and incubated at 55°C for 15 minutes with the phage Lysis Buffer. After the Proteinase K treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 8 cultures using Norgen's Phage DNA Isolation Kit without performing the Proteinase K treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel and the yield of DNA was compared from the eight different phage types (lane 1 to 8). As it can be seen, the optional treatment of Proteinase K improved the phage DNA yield in Lanes 2, 5 and 6 dramatically. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)
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Kit Specifications
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Column Binding Capacity
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50 µg
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Maximum Column Loading Volume
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650 µL
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Size of DNA Purified
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All sizes
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| Maximum Amount of Starting Material |
1 x 1010 pfu/mL enriched phages
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| Average Yield* |
3-15 µg DNA from 108-1010 pfu/mL
of enriched phages |
| Time to Complete 10 Purifications |
45 minutes
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* Average yields will vary depending upon a number conditions used and developmental stage.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 46800 (50 preps) | Cat. 46850 (100 preps) |
|---|---|---|
| Lysis Buffer B | 40 mL | 2 x 40 mL |
| Wash Solution A | 38 mL | 2 x 38 mL |
| Elution Buffer B | 8 mL | 2 x 8 mL |
| Spin Columns | 50 | 100 |
| Collection Tubes | 50 | 100 |
| Elution Tubes (1.7 mL) | 50 | 100 |
| Product Insert | 1 | 1 |
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