I can’t believe I’m already half-way through my Senior Project. Indeed, time flies when you’re having fun.
Over the past two blog posts, I heavily discussed my independent research on the tumor suppressor protein, p53 (go check those blog posts out if you haven’t yet). However, I wanted to spend this week’s blog post specifically talking about my internship and things that have been ongoing in the lab.
For the past few weeks, I’ve been concentrating my efforts in carrying out a procedure known as Restriction Enzyme Digestion. I discussed the intricacies behind this procedure in Week 3’s blog post (I recommend you read this blog first to understand the rest that is to follow). The DNA I’ve been working with over the past few weeks is called pET30-6H-Flu-A-NC. The purpose of this procedure was to observe the quantity and sizes of DNA fragments obtained. However, as all lab procedures sometimes do, I didn’t get the results I necessarily wanted. I could’ve made all sorts of errors:
- Too much buffer solution
- Didn’t pipette any DNA into the Eppendorf tube
- Forgot to add fluorescent dye to the agarose gel
Therefore, prior to re-conducting the Restriction Enzyme Digestion procedure with pET30-6H-Flu-A-NC, I attempted the procedure using a different kind of DNA to ensure that I can perfect it with pET30-6H-Flu-A-NC. Using a DNA called Plu-A-NC, I digested the DNA using Restriction Enzymes NdeI and XhoI. Luckily, the procedure was a success! The results are displayed below:
The lines that traveled further from the top are smaller, while those that traveled less from the top are larger. I know this because smaller fragments of DNA are able to travel through the agarose gel faster than the larger fragments of DNA.
As this procedure was a success, I look forward to successfully digesting pET30-6H-Flu-A-NC with Ecor-I and XhoI next week. Additionally, next week I will go more in depth about the results and what they imply in relation to my research question. Stay tuned!
That’s pretty much all for this week. Thanks for reading, and get ready for next week’s blog post!