Biotech Boot camp and Intro to cDNA Project

    This semester we began lab with a 'biotech boot camp' to both remind returning students and tech new ones about different procedures and protocols for the lab. We covered topics like metric units and conversions, lab notebook etiquette and labeling things. We also went over creating media, using pipettes, experimental design, serial dilution and some basic lab math. 


    At the end of boot camp we went over our new projects, as this semester we will be focusing on Deinococcus papers. I will be working on a project lead by Chad; AMC pathways and qPCR. We went over the basic pathway of the project, a lot of vocabulary and read a few papers covering different techniques we will be covering during our project. 

    

    We will putting D.rad under oxidative stress using Hydrogen peroxide. This will cause the cells to start distribute and receive 'warning signals' called AHL's. 30 minutes after being put in hydrogen peroxide, the cells start to produce enzymes that break down the AHL signals. While most cells only produce one enzyme or the other, D.rad produces both Lactonase and Acylase. 

    At that point I had to go to class, and when I returned they were covering qPCR. qPCR is like regular PCR but measures change in gene expression in real time (called fold change) using a florescent detector. We start with a serial dilution of cDNA (reverse transcribed RNA), in order to get a standard curve (although previous experiments have shown that the best dilution is between 1/1,000 and 1/10,000). For qPCR we do a 10 microliter reaction, 4 microliters of the diluted cDNA and 6 microliters of SYBR Green mastermix. SYBR Green is a dye that binds in-between strands of DNA and is what is measured by the florescent detector during qPCR. 


I would like to include a disclaimer that I know I'm understanding some things incorrectly or not as well as I should, and have large gaps in my knowledge. I will do my best to learn more and understand better both in and out of lab. If you notice something wrong or can describe it in better detail, please leave a comment!


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