Posts Tagged “in gel digestion”

Tips for Sample Preparation in Gel Digestion

Real Experience Q&A

Background

I’m a researcher and I met this problem in my studies. Here it is:

Q: After immunoprecipitation, I obtained protein and made electrophoresis and then in gel digestion for sequencing. But the results showed 80% of keratin. My sequencing teacher said it might be because contamination and I needed to prepare new sample. Can you tell me what do I need to pay attention to in order to avoid contamination? Is keratin contamination very common?

A: Keratin contamination is indeed very common. To avoid contamination, you need to pay attention to the following points:

  1. You in gel digestion process is better to be completed in a ventilated place (such as a fume hood columns);
  2. Try not to do talk when conduct the process and wear hat, masks and gloves;
  3. The glassware to be used should be steeped with 1M HCl for 24 hours and then rinse

The above points should be able to effectively prevent keratin contamination.

Q: Are electrophoresis, staining and digestion also very easy to be contaminated?

A: according to my experience, it’s better to wear gloves, masks and gloves and even bundle the hair