Week 7 – Analyzing RE Digests and Considering Project Direction
Hello everyone,
Restriction enzyme digests are used to cut DNA into fragments. Then, these fragments can be analyzed with gel electrophoresis that separates these fragments by size (smaller fragments traveling farther). This week, I started my project work by looking at my previous gel digests on the vector and “Cxcr3delC” insert (the vector delivers the insert into breast cancer cells). First, I looked at the original PstI digest (Fig. 1) to understand which vector+insert groups I chose to transfect cells with.
Selecting groups 5, 8, 12, 14, and 16, I chose to rerun a restriction enzyme digest after purifying and nanodropping them (processes that clean up the sample to have only DNA remaining). The second gel (Fig. 2) showed cleaner bands alongside a test EcoRI digest that just cuts around the insert. The EcoRI test allows me to check that the vector has taken up the insert (Cxcr3delC) in the first place.
Next week, my mentor is traveling to London so I will not be able to work in the lab. In next week’s blog post, I will discuss the lab work I was able to complete before my mentor left and discuss the specific bands that PstI makes with the vector+insert. Moving forward, I will focus my work more on troubleshooting and bioinformatics. Thank you for reading my blog post and feel free to leave any comments.