Troublesome Site-Directed Mutagenesis: Troubleshooting Your Experiment for Stubborn Mutations

Troublesome Site-Directed Mutagenesis: Troubleshooting Your Experiment for Stubborn Mutations

As is sadly the case in many experiments, site-directed mutagenesis (SDM) does not always work the way we would like it to the first time around. Here are a few tips to help you on your way when trying to troubleshoot a bothersome SDM reaction!

Top Tips for Troubleshooting In Vitro Transcription 

Top Tips for Troubleshooting In Vitro Transcription 

The Phobia of RNases My first experience of troubleshooting in vitro transcription came when I was synthesizing RNA In-Situ Probes for the first time. A lab mate ominously warned me that I had just returned to lab after a bout of flu and that meant I’m a walking talking factory of RNases. I went ahead…

Couples Counselling for Zebrafish: How to Optimize Breeding Efficiency

Couples Counselling for Zebrafish: How to Optimize Breeding Efficiency

It’s Sunday morning, the sun has just begun to rise, and you find yourself on the way to the lab (again!), sipping hot coffee and melancholically thinking of your abandoned bed. But something is different this time. Today, the freezing-cold wind blowing from behind is not the only motivation pushing you to sacrifice another weekend in…

Bacterial Transformation Troubleshooting for Beginners

Bacterial Transformation Troubleshooting for Beginners

The first time I did a transformation was when I worked with site directed mutagenesis. I cloned a protein sequence into the p15TVL vector, created my mutants (but that’s another story), and was finally ready for the next step: transformation and expression of my desired protein. Little did I know that my enthusiasm would fall…

Don’t Let Bubbles Burst Your Experimental Excitement

Don’t Let Bubbles Burst Your Experimental Excitement

Bubbles isn’t just the name of my favorite cartoon character from Power Puff girls, or just the best activity for a kid to play with, in general. In my adult world, they stand for a whole lot more, but can still cause extreme emotions. At the lab bench, seeing bubbles brings happiness or sadness depending…

Are Proteins Adsorbing to Your Labware?

Are Proteins Adsorbing to Your Labware?

One of my favorite things about being a biochemist is to imagine everything at the molecular level—sometimes, in very corny ways. I envision the proteins I pipet and mix as dynamic characters in a molecular soap opera that intermingle with each other in complex ways. The biomolecular characters in my soap opera interact and react,…

Troubleshooting Thin Layer Chromatography:  Some TLC for Your TLC

Troubleshooting Thin Layer Chromatography: Some TLC for Your TLC

The whole TLC technique sounds easy to do, but it can be difficult and tricky during interpretation or give unexpected results, especially when working with biomolecules. For this reason, it is important to be familiar with troubleshooting thin layer chromatography. Some of the common problems faced during TLC and their solutions are listed below: Solvent…

How to Light Up your Life – Tips and Tricks to Troubleshoot your Luciferase Assay.

How to Light Up your Life – Tips and Tricks to Troubleshoot your Luciferase Assay.

What is a luciferase assay and what is it useful for? A luciferase assay takes advantage of the innate bioluminescent properties some organisms exhibit, most notably the firefly. The firefly can convert luciferin to oxyluciferin in the presence of the enzyme luciferase to emit light. The most common scientific assays utilizing luciferase are reporter assays…

Troubleshooting: No Events on Your Cytometer

Troubleshooting: No Events on Your Cytometer

It’s happened to us all, you are ready to run your samples on the cytometer and you can’t see your cells on the screen. Here are a few tricks to troubleshooting this: Cytometer vs. computer connection The different types of cytometers will need different orders for switching on the cytometer and computer. Some are cytometer…

How to Troubleshoot Problems with Fluorescently Tagged Proteins
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How to Troubleshoot Problems with Fluorescently Tagged Proteins

Using fluorescent proteins as imaging probes is a widespread and versatile technique in microscopy. You can use them in a wide range of living systems, from single cultured cells to complete organisms and animals. Fluorescently tagged proteins can be used to track and examine real-time localization, interactions and translocation of your protein of interest, as…

Garbage in, Garbage out? Quality Control of Your NGS Data

Garbage in, Garbage out? Quality Control of Your NGS Data

So, you’ve just received a call from the core facility that you hired to prepare and sequence your libraries. The facility director tells you that the sequence data from your next generation sequencing (NGS) experiment does not look good. You panic and, perhaps, let loose a scream of frustration—aaarrrrggghhhh! This project was going to be…