Key Tips to Overcoming Experimental Hurdles

May 10, 2017

No matter how much research experience you have, you can encounter roadblocks with your experiments at any time. You may be creating a mutant strain of bacteria, purifying proteins, or using flow cytometry and hit a hurdle with the experiment. What do you do next? We provide key tips on overcoming hurdles with experiments in the lab.

Choose Your Controls Wisely

The number one thing you learn while conducting research is using controls. Remember to think about each step of the protocol and the correct controls to set up so that you can determine whether the protocol works. The best way to discern this is to use a positive control - a factor that you know works if the protocol was done correctly. For example, if you are separating a protein by its cellular location (i.e., inner or outer membrane), then also assess proteins you know that reside in the inner and outer membranes to determine the effectiveness of the protocol.

If you've done this and discovered that the positive control is not working, then you know something is wrong with the protocol or with the materials. Maybe the restriction enzyme isn't working anymore, or the buffers were made incorrectly. Which brings us to our next point.

Be Meticulous About Protocols

"Sometimes little things can make a big difference. Go over your protocol several times in advance and follow every detail. When making buffers and reagents, make sure they are at the correct pH and concentration. Check reagent stocks, lot numbers, and expiration dates. With kit-based assays, know the function of 'Buffer A' before attempting your experiment. Don't hesitate to call the manufacturer with product-related questions," comments Dr. Alan Goggins, a recent doctoral graduate in the Department of Microbiology and Immunology at Tulane University.

Ask Your Colleagues for Troubleshooting Tips

You learn the most common protocols of your lab from the people that work there. It is only natural to ask them how they dealt with an issue in a protocol. Chances are they ran into the same issues you are experiencing and/or successfully ran the experiment. Dr. Goggins adds, "Don't be shy about reaching out to scientists from other departments or institutions. There is a wealth of online resources and forums where you can post questions and get answers from other scientists."

Talk to Your Principal Investigator (PI) for Their Advice 

Dr. Goggins recommended that you talk to your PI about your experimental struggles, because they have a wealth of experience and knowledge and can likely help you troubleshoot any experiments. "Ph.D. students are hesitant to talk about their failed experiments with their PIs because they feel it reflects poorly on their scientific abilities, but your mentor is there to help," commented Dr. Goggins.

Think About the Big Picture of Your Research

Dr. Goggins said, "Sometimes experiments fail for reasons beyond your control. Think hard about the question an assay is trying to answer and whether it's actually the correct application. Try to think about alternative experiments or assays that could be applied to test your hypothesis. Think about your experimental results within the context of the overall hypothesis. Remember that there's a big difference between a failed experiment and negative data."

Work on Something Else Simultaneously 

The satisfaction of having another assay work will keep you going as you troubleshoot the problem with the another experiment. Floricel Gonzalez, a Ph.D. student in the biological sciences program at Virginia Tech and Undergraduate Research Fellowship Alum agreed. "Recently, I was pioneering a new assay in the lab and everything was going smoothly. I was halfway to generating the data for the complete experiment set when the assay stopped working. I retried the experiment a couple more times and had the same confusing results. This is when I tested every reagent and found that one of my stocks was contaminated. Once I filtered this stock, I was able to fix the problem. As frustrating as this setback was, I had other experiments going on that were working and that motivated me to keep pushing forward."

Mistakes Happen: Learn From Them

Sometimes a "no-duh" type of mistake happens. Don't beat yourself up, or think any less of yourself as a researcher. Everyone makes mistakes at any job. The most important thing is to learn from your mistakes and move forward. Dr. Goggins added, "The great thing about failing is that failure is often the best opportunity for growth and learning."

With these tips, we hope that you can overcome obstacles with experiments in the lab!

Author: Shilpa Gadwal

Shilpa Gadwal
Shilpa Gadwal is the Career Resources Specialist at ASM.