Bacteriophage Therapy Unleashed: From Theory to Lab Practice

Every great experiment starts with the right tools. In this practical guide, you’ll learn how to prepare LB, TSB, and soft agar; make SM buffer and PBS; and sterilize equipment safely. We’ll walk through essential lab supplies, explain why each reagent matters, and share pro tips to avoid contamination. Whether you're setting up a teaching lab or starting independent research, this lecture gives you the foundation for success.

The secret to successful phage experiments? Healthy, actively growing bacteria. In this step-by-step tutorial, you’ll learn how to grow bacterial cultures to mid-log phase — the optimal window for phage infection. We’ll cover OD₆₀₀ measurement, CFU/mL estimation, incubation conditions, and common pitfalls. After this lecture, you’ll never guess when to infect again — you’ll know exactly.

Found a sample with potential? Now boost your chances of finding phages inside. In this lab-focused lecture, you’ll master enrichment protocols using host bacteria to amplify low-abundance phages from sewage, feces, soil, or wound swabs. We’ll cover incubation times, temperature, and troubleshooting tips to maximize recovery — turning weak signals into strong, detectable lysates.

The spot test is your first line of defense in phage detection. In this fast-paced, demo-driven lecture, you’ll learn how to perform a reliable spot test — from preparing the lawn to interpreting clearing zones. We’ll show you how to distinguish true lysis from inhibition, optimize spotting volume, and troubleshoot false negatives. It’s quick, simple, and essential for every phage hunter.

Want to watch phages kill bacteria in real time? The turbidity reduction assay lets you do exactly that. In this detailed walkthrough, you’ll learn how to set up the assay, calculate MOI, track OD₆₀₀ over time, and interpret lysis curves. Whether you're screening phages or comparing efficacy, this quantitative method is a must-have tool in your arsenal

No soft agar? No problem. In this innovative lecture, you’ll learn the reduction streaking method — a powerful alternative to double agar overlay for isolating phages. Using sterile loops and quadrant streaking, you’ll progressively dilute phages until single plaques appear. It’s perfect for labs with limited resources or when working with fast-lysing phages.

The gold standard for phage enumeration. In this comprehensive lecture, you’ll master the double agar overlay (DAO) technique — from molten soft agar handling to plaque counting. We’ll cover dilution strategies, ideal plaque density, and how to calculate PFU/mL with confidence. By the end, you'll be able to titer any phage stock like an experienced researcher.

A single plaque doesn't guarantee purity — unless you do it right. In this meticulous lecture, you’ll learn how to pick, elute, and purify a single plaque — and repeat the process three times to ensure clonality. We’ll show you best practices for avoiding contamination, recognizing uniform morphology, and confirming a pure isolate. This is where amateur work ends — and professional phage science begins.

Fecal samples can contain both phages AND antibiotic residues — and confusing them can ruin your experiment. In this critical lecture, you’ll learn diagnostic tricks to differentiate between the two: heat sensitivity tests, filtration checks, DNase treatment, and host-specificity assays. This skill is vital for gut microbiome studies and clinical phage isolation projects.

Get up close and personal with the virus you’ve been studying. In this awe-inspiring lecture, we explore how electron microscopy reveals phage morphology in stunning detail. You’ll learn sample preparation, staining techniques (negative stain), and how to classify phages into Myoviridae, Siphoviridae, and Podoviridae based on tail structure. Even if you don’t have access to a TEM, understanding these images is crucial for publication and identification.

One phage goes in — how many come out? That’s the burst size, a key measure of phage fitness. In this calculation-heavy but accessible lecture, you’ll learn how to derive burst size from your one-step growth curve. We’ll walk through the math, explain what high vs low burst sizes mean, and show how this impacts therapeutic potential.

When does a phage go from silent invader to explosive replicator? The latent period holds the answer. In this advanced lecture, you’ll design and interpret a one-step growth curve — measuring the eclipse and burst phases. We’ll guide you through infection synchronization, plating, and data analysis so you can determine exactly how fast your phage replicates.

Is your phage a T4-like podovirus or a lambda-type siphovirus? In this molecular biology deep dive, you’ll learn how to extract phage DNA and use PCR with specific primers (e.g., g20, terL) to identify conserved genes. We’ll cover primer selection, degenerate bases, and BLAST analysis — turning your unknown phage into a named, classifiable agent.

Your hard work deserves protection. In this essential final lecture, you’ll learn how to store phage stocks long-term without losing viability. We’ll compare refrigeration, freezing, and ultra-cold storage; demonstrate glycerol preservation; and teach you how to avoid freeze-thaw damage. Plus: chloroform use, aliquoting, labeling, and revival techniques — so your phages survive for months or even years.