While we all love to do gene expression analysis of our treated cells, there is always one thing we need to do before any of our data can be published: primer validation. This task is what our 6-week internship student Sophie Schwitzgoebel took care of.
Before performing an actual PCR, Sophie blasted all of our primer pairs online to check if they align with the respective, intended genes. This was followed by in-silico PCR analysis to check if the primer pairs amplify the correct gene. She performed PCR to check for the correct size of the amplicon and sent her samples off for sequencing.
Besides doing a great job in characterizing all of our primers, Sophie also set up a fantastic poster which summarizes the most important facts all around primers. Not only does she describe how to check for in-silico primer specificity, but she also noted why it is important to prioritize the usage of exon-spanning primers. Of course, these tests should be done during the design process, so she also included a very nice description of the most important points to look for when designing new primers. We really enjoyed the poster presentation :).
She also summarized her most important in-silico validation and sequencing results in a very nice and structured table which you can find below.
We thoroughly enjoyed having Sophie as an internship student and are grateful for her important work of testing all of our primers for specificity, thereby ensuring that we only publish verified data sets. We wish you all the best with your upcoming master thesis in Leipzig!
Hannah
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