Why is our gel electrophoresis gel sticking to the sides and ripping?

  • Thread starter jesswess
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In summary, a biophysics grad student at Michigan State is having trouble with agarose gels sticking to the sides of the form and ripping, rendering them unusable. The protocol being followed has worked for the PI in other labs, but not in this new lab with new equipment. The student has ruled out contamination and has tried suggestions such as letting the gel sit longer or putting it in the fridge. They are seeking advice on what could be causing the issue and are considering contacting the company for help.
  • #1
jesswess
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I am a biophysics grad student at Michigan State, and the lab I am working in has just started making 1% agarose gels for DNA purification. The problem is when we try and remove the comb from the gel and the gel from the walls of the form, it sticks to the sides, ripping the gel and rendering it unusable. The protocol that we are following as worked for my PI in other labs, but never for myself (or my PI) in this lab. All our equipment, chemicals, and glassware are new because this is a new lab.

So, have any of you had a similar problem? What do you think could be causing this?

I have made sure that it is not the agarose or TBE buffer that is contaminated.
Here is the protocol:
Add 0.6g of low melting point LMP Agarose (UltraPure brand) to 60mL of 0.5x TBE buffer (Fisher brand). Microwave until just boiling and mix until all the agarose is dissolved. Allow to cool, add 1μL of EtBr and pour into mold. Wait ~1 hour and remove gel from mold.

Like I said, my PI used this protocol all the time at his past lab, and he never had this problem.
Thanks so much for your help.
 
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  • #2
Have you tried letting it sit for a little longer? The only other thing I can think of is putting it in the fridge for the last 15 min. or so.
 
  • #3
I made hundreds of 1% gels, also on new equipment, and have never had that happen. Be gentle when removing the comb, it helps to start the lifting from one side so that air can come into it. Otherwise contact the company for advice.
 

1. What is gel electrophoresis?

Gel electrophoresis is a laboratory technique used to separate and analyze DNA, RNA, or proteins based on their size and charge. It involves running a sample through a gel matrix using an electrical current, causing the molecules to move towards the positive electrode at different rates.

2. Why is gel electrophoresis important?

Gel electrophoresis is an important tool in molecular biology and genetics research. It allows scientists to separate and visualize different molecules, such as DNA fragments, which can provide valuable information about genetic variation and relationships between organisms.

3. How do I prepare a gel for electrophoresis?

To prepare a gel for electrophoresis, you will need to mix a specific concentration of agarose or polyacrylamide powder with a buffer solution and heat it until it forms a gel. Once the gel has solidified, you can create wells for loading your samples and then run the gel using an electrical current.

4. What factors can affect the results of gel electrophoresis?

Several factors can affect the results of gel electrophoresis, including the concentration and type of gel used, the voltage and duration of the electrical current, and the size and type of molecules being separated. Contaminants or errors in sample preparation can also impact the results.

5. How can I analyze the results of gel electrophoresis?

The results of gel electrophoresis can be analyzed by visualizing the gel under UV light and comparing the bands or peaks of the separated molecules to known standards or other samples. Software programs are also available to help quantify and analyze the results.

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