jmuhling

I run the GSEx viral vector core at the University of Adelaide as my own personal fiefdom. My favourite activities are snacking and making fun of graduate students. I will talk about science to anyone who wanders within range.

Articles by jmuhling:

The Three Ts of Introducing Foreign DNA: Transfection, Transduction, and Transformation

Introducing foreign DNA into bacterial or eukaryotic cells is a common molecular biology technique. Some of the terminology involved can be confusing: transformation, transfection and transduction. What do these words actually mean? To make matters worse, sometimes people use these terms interchangeably, resulting in confusion between colleagues and collaborating labs. Here then is your definitive guide…

24 Jan 2017 DNA / RNA Manipulation and Analysis

Outsourcing Research: Should Your Experiment Spend Some Time Away from You?

As a researcher, it’s satisfying to manage your own projects and do the bench work yourself. After all, if you don’t have experience with a technique, you’re usually expected to figure it out (with or without direct supervision). In some situations, dealing with difficult molecular techniques is simply part of the job description. The scientific…

18 Jan 2017 Basic Lab Skills & Know-how

Viral Vector Production: Myths & Misconceptions

Viral vector production is a worthwhile skill that can be made even easier with a few tips and tricks. In general, transfection of multiple plasmids into a producer cell line results in infectious, non-replicative virus. However, it is important to ensure that your vector preparation is efficient, giving your experiments the best chance of success.…

10 Jan 2017 DNA / RNA Manipulation and Analysis

AAV Production Part II: Virus Purification

In Part I of AAV Production, I described how to produce crude (non-purified) AAV. In this article, I am going to tell you how to purify that crude prep. Virus purification is usually done by gradient ultracentrifugation. Two common methods involve gradients made from increasing concentrations of cesium chloride or iodixanol. A cesium chloride prep…

13 Dec 2016 DNA / RNA Manipulation and Analysis

Why You Should Defend Science

It’s probably safe to say that most people reading this article are big fans of science. As scientists, we love using rigorous methods to learn more about how the world works. So it may come as a shock to realize that science is often viewed as flawed, immoral or just plain wrong – especially when…

30 Nov 2016 Career Development & Networking

AAV Production Part I: Tips for Setting Up

What is Adeno-Associated Virus? Adeno-associated virus (AAV) is a popular gene therapy vector. Different AAV serotypes infect different cell and tissue types with varying efficiency, so it’s a good idea to select the serotype most relevant to your work. The most commonly used AAV serotypes are 1, 2, 5, 6, 8, 9 and DJ, although…

15 Nov 2016 Cells and Model Organisms

How to Effectively Organize a Research Lab

An academic lab is a unique working environment. Lab members are expected to take responsibility for their own research projects and perform the work quickly and efficiently. However, unlike an industrial or corporate setting, there are often no clearly defined management structures. This means that when it comes to communal equipment, reagents and resources, individual…

17 Aug 2016 Organization & Productivity

How to Best Improve Your Lentivirus Titer

If you’re planning on using lentivirus for gene therapy, you will need to find out how many lentivirus particles to use. However, this is where researchers new to gene therapy work often run into problems. If this is you. Stop. Before you embark on endless optimisation experiments (i.e. a different transfection reagent, new cells, different phases…

09 Jul 2016 Protein Expression & Analysis

Want to Increase Your Lentiviral Titers? Focus on Your 293T Cells

Producing lentiviral or retroviral vectors is theoretically fairly straightforward. However, anyone new to viral vector work is usually confronted with vast amounts of confusing information. It seems that anyone who has ever made a lentivirus has their own protocols and is adamant that their method is the best one to follow. In reality, there are…

09 Jul 2016 Cells and Model Organisms

How (not) to write a terrible scientific thesis

So it’s time to write up your thesis. By now, you probably consider yourself an expert on your topic or you hate the sight of it (or both!). Either way, hundreds of typewritten pages are all that stands between you and graduation/freedom. One of your first requirements will be to review the current knowledge on…

09 Jul 2016 Writing, Publishing & Presenting