DNA purification from bacteriophages

Norgen Biotek's Phage DNA Extraction Kit provides fast and efficient purification of total DNA from bacteriophages.

Spin column

Size

• 50 Preps
• 100 Preps

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Image Credit: Norgen Biotek Corporation

Image Credit: Norgen Biotek Corporation

Image Credit: Norgen Biotek Corporation

Image Credit: Norgen Biotek Corporation

Overview

Features and benefits

• Extract superior DNA from a wide range of phage strains.
• High amounts of total DNA
• Processing in a rapid spin-column format is quick and simple
• High yields of DNA were recovered without the need for cesium chloride banding or phenol or chloroform extractions.
• 3–15 µg of enriched phage DNA from 10⁸–10¹⁰ pfu/mL

This kit utilizes a rapid spin column method to purify total DNA from a wide range of bacteriophages cultured in liquid-grown bacteria. Phenol, chloroform, or cesium chloride banding techniques are not used in the isolation of the DNA. In less than 45 minutes, the spin-column-based process can be finished.

With the optional DNase and Proteinase K treatments, phage DNA yields are increased while host DNA contamination is reduced, and the kit is very effective at processing small volumes of phage supernatant (500 µL to 1 mL). PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot, and other downstream processes can all use purified total phage DNA, which has the highest integrity.

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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 1× 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 DNase I efficiently degraded the host genomic DNA. 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). Image Credit: Norgen Biotek Corporation

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). Image Credit: Norgen Biotek Corporation

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Kit specifications

Source: Norgen Biotek Corporation

* Average yields will vary depending upon a number conditions used and developmental stage.

Storage conditions and product stability

Every solution needs to be stored at room temperature and kept tightly sealed. This kit remains stable for one year from the shipment date.

Source: Norgen Biotek Corporation