Phage DNA Isolation Kit

Features and Benefits

Isolate high quality DNA from a broad variety of phage strains
High yields of total DNA
Fast and easy processing using a rapid spin-column format
No phenol or chloroform extractions or cesium chloride banding required
High yields of DNA recovered 3-15 µg DNA from 10⁸-10¹⁰pfu/ mL of enriched phages
Phage whole genome sequencing services available

This kit provides a rapid spin column method for the purification of total DNA from a broad spectrum of bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL) and with the optional DNase and Proteinase K treatments phage DNA yields are maximized while host DNA contamination is minimized. Purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot and more.

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)

Kit Specifications
Column Binding Capacity	50 µg
Maximum Column Loading Volume	650 µL
Size of DNA Purified	All sizes
Maximum Amount of Starting Material	1 x 10¹⁰ pfu/mL enriched phages
Average Yield*	3-15 µg DNA from 10⁸-10¹⁰ pfu/mL of enriched phages
Time to Complete 10 Purifications	45 minutes

* 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

Documentation

FAQs

Spin Column

I noticed the column was clogged. What causes this?

Column clogging may occur due to one or more of the following reasons:

Centrifugation speed was too low or spin time was inadequate.
Check the centrifuge to ensure that it is capable of generating the required RPMs. Sufficient centrifugal force is required to move the liquid through the resin. Also, ensure that the correct spin times are followed. Spin for an additional minute if necessary.
Bacterial debris in the lysate.
Ensure that the starting material is clarified phage supernatant. Remove bacterial debris from the initial phage supernatant by centrifugation at 10,000 × g for 5 minutes before beginning the protocol.
The lysate/binding solution mixture is not homogeneous.
To ensure a homogeneous solution, vortex for 10-15 seconds before applying the lysate to the spin column.
Centrifuge temperature is too low.
Ensure that the centrifuge remains at room temperature throughout the procedure. Temperatures below 20℃ may cause precipitates to form that can cause the columns to clog.

Why do I have low yield of phage DNA?

The yield of genomic DNA may be lower than expected due to the following:

Ineffective propagation of phage and initial lysis step.
Refer to the manufacturer's recommendations for the propagation of the phage, including proper titer for inoculation, growth conditions, and bacterial host.
Incomplete lysis of cells.
Ensure that the incubation was performed for 15 minutes at 65℃ after the addition of Lysis Buffer B. Also, perform optional digestion with Proteinase K in Step 1b during Lysate Preparation.
Ethanol was not added to the Wash Solution A.
Ensure that 90 mL of 96 - 100% ethanol is added to the supplied Wash Solution A prior to use.
The DNA elution is incomplete.
Ensure that all the DNA is eluted. If elution buffer remains in the column, use 14,000 g for the second centrifuge.

Why is the purified DNA not performing well in downstream applications?

If the DNA does not perform well in downstream applications, it may be due to one or more of the following:

DNA was not washed three times with the provided Wash Solution A.
Traces of salt from the binding step may remain in the sample if the column is not washed three times with the Wash Solution A. Salt may interfere with downstream applications and thus must be washed from the column.
Ethanol carryover.
Ensure that the dry spin under the Column Wash procedure is performed in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.

Why do I have host genomic DNA contamination in phage DNA elution?

Host genomic DNA can contaminate the phage DNA. In order to eliminate host genomic DNA contamination in the phage DNA elution, it is recommended that a DNase I treatment is performed at the beginning of the lysate preparation procedure (see Section 1, Optional DNase Treatment).

Can this kit isolate phage genomes with ssDNA ?

Yes, the kit can isolate both ssDNA and dsDNA phage genomes.

Can PEG precipitation be used before the Phage DNA isolation kit?

The kit doesn't require PEG precipitation. However, the kit is compatible with PEG precipitated samples.

Title	Isolation and characterisation of a novel Stenotrophomonas maltophilia phage vB_SmaS_BCU-1 with evaluation of mammalian cell safety
Citation	European Journal of Clinical Microbiology & Infectious Diseases 2026.
Authors	Kashif Haq, Martin Figgitt, David Lee, Jack Spencer & Anisa Choudhry

Title	Timely bespoke phage-antibiotic combination to treat refractory Pseudomonas aeruginosa mediastinitis and vascular graft infection
Citation	Nature Communications 2026.
Authors	Shimin Jasmine Chung, Yang Liu, Shuhua Thong, Yang Zhong, Zhi Soon Chong, Zhining Lim, Sabrina Tan, Jia Hao Yeo, Ming Guang Koh, Nathalie Grace Sy Chua, Dorothy Hui Lin Ng, Winnie Hui-Ling Lee, Tze Peng Lim, Limin Wijaya, Boon Huan Tan, Peng Huat Eric Yap, Thet Tun Aung, Rick Twee-Hee Ong, Karrie Kwan Ki Ko, Tse Hua Nicholas Wong, Yu Lin Charlene Tang, Yee Jim Loh, Teing Ee Tan, Thuan Tong Tan, Andrea Lay-Hoon Kwa

Title	Functional mechanism and clinical implications of lncRNA NNT-AS1 in delayed fracture healing
Citation	Cytotechnology 2026.
Authors	Phil Huss, Chutikarn Chitboonthavisuk, Anthony Meger, Kyle Nishikawa, R. P. Oates, Heath Mills, Olivia Holzhaus, Srivatsan Raman

Title	Isolation, Identification, and In Vivo Anti-Infective Efficacy of Lytic Bacteriophage SEP1 Against Salmonella Paratyphi
Citation	BMC Microbiology 2026.
Authors	Zhiyi Ge, Di Lian, Wei Zhao, Weiru Song, Shengyi Han, Chunyan Xu

Title	Electrospun Phage-Loaded Bilayer Nanofibrous Scaffolds for Wound Dressing Applications: A Comparative Study of Different Bacteriophages
Citation	Journal of Functional Biomaterials 2026.
Authors	Siavash Aghili, Muhammed Awad, Md Hasib Adnan, George Bouras, Tran Thanh Tung, Sarah Vreugde and Dusan Losic

Title	Three Staphylococcus Bacteriophages Isolated from Swine Farm Environment in Quebec, Canada, Infecting S. chromogenes
Citation	Viruses 2026.
Authors	Mousumi Sarker Chhanda, Rébecca E. St-Laurent, Valérie E. Paquet, Nicolas Deslauriers, Cynthia Gagné-Thivierge, Martine Denicourt, Marie-Ève Lambert, Antony T. Vincent, and Steve J. Charette

Title	Isolation and characterization of bacteriophages with lytic activity against multidrug-resistant non-typhoidal Salmonella from Nairobi City county, Kenya
Citation	BMC Infectious Diseases 2025.
Authors	Michael Mugo, Abednego Musyoki, Angela Makumi, Ivy Mutai, Kelvin Kering, Peter Muturi, Collins Kebenei, Kristin Weber, Michael Pietsch, Tanja Pilz, Oliver Drechsel, Tobias Hoffmann, Lothar Wieler, Cecilia Mbae, Antje Flieger & Samuel Kariuki

Title	Characterization and application of a novel Campylobacter phage CC_R7 as a biocontrol agent in chicken meat
Citation	Current Research in Food Science 2025.
Authors	Muhammad Shahzad Rafiq, Jie Chen, Muhammad Akmal, Dongxin Ma, Ali Asif, Junhao Wang, Shuaifeng Gu, Yufeng Gu, Pan Tao, Haihong Hao

Title	Characterization and therapeutic evaluation of the lytic bacteriophage ENP2309 against vancomycin-resistant Enterococcus faecalis infections in a mice model
Citation	Virology Journal 2025.
Authors	Jiaqi Tian, Luyao Wang, Rui Gao, Wenwen Zhou, Shinan Zhang, Lingxia Li, Guoyuan Hu, Licheng Xiao, Yijuan Ma, Sang Ba, Daijiyongzang, Shengyi Han, Shengqing Li

Title	Isolation and Characterization of Four New Coliphages against Extra Intestinal Pathogenic Escherichia coli Isolates Recovered from Cystic Fibrosis Patients
Citation	ACS Omega 2025.
Authors	Gerardo García-González, José Manuel García-Pérez, Patricia Dolores Martinez Flores, Ruth Rivera Ríos, María del Rosario Espinoza-Mellado, Norma Velázquez-Guadarrama, David Armando Encinas Basurto, Josué Juárez, Marco Antonio López Mata, Carlos Alberto Eslava-Campos, Gerardo E. Rodea

Features and Benefits

Details

Supporting Data

Documentation

PROTOCOLS

SAFETY DATA SHEETS

FLYERS

FAQs

Spin Column

I noticed the column was clogged. What causes this?

Why do I have low yield of phage DNA?

Why is the purified DNA not performing well in downstream applications?

Why do I have host genomic DNA contamination in phage DNA elution?

Can this kit isolate phage genomes with ssDNA ?

Can PEG precipitation be used before the Phage DNA isolation kit?

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Phage DNA Isolation Kit