Peddie 2016 Summer Blog Link

http://peddieschoolexp2016.blogspot.com/2016/05/

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Stephanie Wu | Week 3 | More Slicing and Mounting

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06.19.17 - 06.25.17 For the majority of this week I spent my time slicing and mounting the brains Felicia had previously removed from some mice via perfusion after inserting the injections she was working with. The main type of mouse I have worked with are the rAAV2 Delta-Cre mice (which are the controls) and the rAAV2 GFP-Cre mice (which are the experimentals), which are then divided into hetero and homo. From there, Felicia targeted specific regions of the brain, mostly injecting into areas such as the nucleus accumbens (NAc) and the dorsal striatum (DS). She injected both a control virus as well as multiple experimental viruses. By slicing the brains we would eventually be able to image them on the microscope and see the effects of the injection on the synapses of the brains and the different afferent sites as well. (some brain slices (N1C. 87)) (more slices) (some mounted slides of N1C. 125 & 126 brain slices) (what my slide box currently looks like) Bes...
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Emily Guo, Week 7, Hanging Up the Lab Coat

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Some good news: the plasmids Anna and I made a few weeks ago were sent off for sequencing, and they came back with the correct sequence! The plasmids can now be used in the next round of electroporation.  We are currently still troubleshooting the bisulfite conversion process. I ran a PCR for the DNA samples (non-converted) that we were using on Monday with two sets of primers that we knew would work, and we did indeed have bands of DNA show up on the gel afterwards. Therefore, the quality of the DNA that we were converting wasn’t the problem. I found a bisulfite conversion guide by Zymo Research – which deals a lot in epigenetic research in addition to providing biology research tools for DNA and RNA research and analysis – that provided some tips that we could try out. One was a way to analyze the DNA after bisulfite conversion, which we didn’t do before, by running the gel with the bisulfite-converted product. However, because the DNA becomes so damaged through the conversi...
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Kabir Grewal, UPitt Lab, End of Week 4

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I ended my last blog post with writing about when I injected TMSC’s and CSSC’s into  well plates with  the extracellular matrix of bovine corneal endothelial cells (BCE cells) , which brings me to the work I completed last Thursday and Friday. Last Thursday was a busy day for me.  After arriving at the lab at the usual time of 9:30, I checked the growth of the newly injected stem cells mixed with the extracellular matrix of lysed BCE cells. As expected, the stem cells had not attached to the bottom of the well plates, and were free floating in the DF10 growth medium. Today, my task was to make a new growth medium for these cells, as well as create the variables for the experiment.  The new growth medium I made comprised of a glucose-rich base, fetal bovine serum (FBS), as well as ROCK inhibitor (short for Rho Kinase inhibitor). The purpose of this inhibitor is to prevent excessive apoptosis among the cells, and promote the cells to grow and retain their struc...
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