Friday, May 31, 2013

Chelsea McCurry's Internship, Summer 2013 - Part 5

This past week we have worked on preparing three additional sediment samples from the Gulf Sediment and will be shipping them for DNA sequencing next week. We continue to evaluate the sequences we received last week. So far, I have done the preparation of each of the sequences with a computer program. I took out the parts of the sequence that are from the vector we use, trimmed the ends that have a lower quality than the rest of the sequence, and removed any sequences that were too low of a quality for us to use. We can now begin analyzing this data and compare it to the rest of the data we already have.

The picture shows a Petri dish with LB Agar that has X-gal allowing the Blue/White differentiation. The white colonies an E. coli cell that we were able to transform and clone to carry a vector that a single piece of Foraminifera DNA (about 750 bp). This allows us to take the mass amount of DNA we have from a basic extraction and PCR, and isolate and amplify each strand within an E. coli colony. The blue colonies are the E. coli that did not take up any DNA. This is important to be able to differentiate, because we don’t want to spend time and money in sequencing when there is no target DNA there. The next image shows me picking the white colonies and placing one colony into its own well of LB broth in a 96-well plate. I will incubate the samples 12 hours then prepare them to be shipped off for sequencing.  

Posted by:
Chelsea McCurry, University of West Florida

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