Preprint Technical Note Version 1 Preserved in Portico This version is not peer-reviewed

A Modified High-Throughput Screening Protocol to Isolate Bacteriophages from Environmental Samples

Version 1 : Received: 16 February 2021 / Approved: 18 February 2021 / Online: 18 February 2021 (10:46:00 CET)
Version 2 : Received: 18 February 2021 / Approved: 19 February 2021 / Online: 19 February 2021 (09:53:32 CET)

How to cite: Abdelsattar, A.; El-Shibiny, A. A Modified High-Throughput Screening Protocol to Isolate Bacteriophages from Environmental Samples. Preprints 2021, 2021020410 (doi: 10.20944/preprints202102.0410.v1). Abdelsattar, A.; El-Shibiny, A. A Modified High-Throughput Screening Protocol to Isolate Bacteriophages from Environmental Samples. Preprints 2021, 2021020410 (doi: 10.20944/preprints202102.0410.v1).

Abstract

In the post antimicrobial era, increasing attention is paid towards using bacteriophage (phage in short) therapy to control antibiotic-resistant bacteria. The first step in phage therapy applications is isolating highly efficient lytic phages or phage cocktails from various sources. When a double-layer- agar with around 0.7% agar in top agar is employed, it results in a low number of phage isolation with a poor resolution, and in many cases, you miss the phage. To address this problem, a low concentration of agar in top agar is examined for better phage isolation. Here, our results proved the efficiency of isolating phage upon formulating a double-layer agar with 0.3% agar in top agar. A sewage sample was collected then phages were isolated, purified, and spotted on a top layer agar with 0.3% agar. The results showed the possibility of isolating a higher number of phages on 0.3% top agar than 0.7%. The finding advocates using 0.3% top agar for the double-layer agar, as it will provide fast, better, and easy phage screening and isolation.

Subject Areas

Phage Purification; phage isolation; phage characterization; jumbo bacteriophages; top agar; double agar overlay; high throughput sequencing

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