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This post was contributed by guest blogger, Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology.

If you are reading this article then you probably suspect mycoplasma contamination in your cell culture or you are about to begin a new cell culture project. If mycoplasmas are present in your lab, don’t be surprised. They are present in most of the cell culture facilities, tissue culture labs and every cell culturist has to deal with this problem. It is estimated that mycoplasma is responsible for up to 60% of the cell culture contamination (Uphoff, 2002).

Mycoplasmas are considered to be one of the simplest and smallest bacteria. The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial drugs like penicillin and streptomycin. Mycoplasma can pass through filtration methods because of its ability to change shape and the absence of a rigid cell wall. Here, I will cover some of the best ways to tackle mycoplasma contamination before they enter your cell culture and what to do if you encounter mycoplasma contamination.

One of the main tenets of our quality control (QC) mantra is to do our utmost to ensure that scientists are receiving the exact materials that they think they are. To address this, several years ago we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform that allows us to provide scientists with complete plasmid sequences. After launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed viral genome sequencing (VGS), to confirm the identity and serotype of our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene Therapy.

Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the physical titer of our preps. We often repeat this assay to confirm that our results are accurate. But what if we could repeat this assay 10,000 times to ensure that our titers were as accurate as possible? This is where droplet digital PCR (ddPCR) comes in.

Understanding the biology behind brain development, learning, and behavior poses many challenges in neuroscience with many brain regions, neuronal networks, and specific neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists have begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases, and more. With the help of adeno-associated viral vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation of specific neuronal subtypes and determine how these changes affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed so there are many combinations to choose from for your experiments!

Dear scientists using AAV,

We hear your questions! How much virus should I inject? How long do I need to wait? What serotype and promoter should I use to target my favorite neurons?

We don’t know all of the answers, but we’re betting that your colleagues do. That’s why we’re launching the AAV Data Hub. The Data Hub is an open platform for scientists using AAV to share practical information about their experiences. It will contain all of the juicy and important details that you won’t find in the final publication and we have already collected over 100 entries spanning six different species and dozens of different expression sites.