Post 5: Final Thoughts about the Summer

Hello again!

Well, the summer is officially over. I’ve put in 10 long weeks of work and I’ve accomplished quite a bit. While this series of blogs has been focused on my honors thesis project, I’ve also been working on the tRFLP project described in last summer’s blog series. With that project, I have gotten fragment data for every month from Mat 2009 to August 2010! The next step is the analysis of the fragments and creation of dendrograms to compare similarity in the bacterial community in each month over the course of the year. I’ve got my work cut out for me there too!

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Post 4: Mini Prep Mayhem

Hi All,

To recap – I began the summer by extracting bacterial DNA from 0.22 micron filters that held bacteria from Lake Matoaka. I then amplified the 16s rRNA gene by PCR. These amplicons were ligated into plasmid vectors, transformed into E. coli cells and hundreds of copies of each amplicon were made. What’s next? Since the end goal is to sequence the bacterial 16s gene, I need to get the plasmid back out of the E. coli cell. Enter: Mini-Prep kits. These are brilliant little kits that have made the process of removing plasmid DNA much easier.

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Post 3: Transformation Nightmare!!

Hi All,

You’re probably dying to know the results of the transformation reactions that I set up last post! Sorry it’s taken me so long to get back to you all!

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Post 2: Ligation and Transformation

Hello All!!

After several weeks of DNA extractions, I finally moved on to the next steps in my thesis research!!

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Post 1: DNA Extraction

Hi All,

I realize that June is almost over and I haven’t posted any updates about my research. June has been a very busy month for me.  But, the bulk of my work has centered around the extraction of bacterial DNA from the 0.22 micron filters that are used to collect the bacteria from Lake Matoaka on a monthly basis. So far this month, I’ve extracted nine months (and counting) worth of DNA from filters that have been stored at -80 C for almost a year now. 

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