Week 1- PCR!

Another project that I was able to work on during my first week was PCR using DNA from different samples of ergot, a fungi that grows on plants such as blue grass and wheat. If ergot is ingested it can lead to vasoconstriction and can also cause hallucinations.

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Different samples of ergot DNA

To perform the PCR, we had to combine primers, water, and master mix into each capsule along with the different DNAs. The master mix is a bright green color because it contains ingredients that help it sink into the gel later in the PCR. 11652138_737365056372926_958455970_nWhen pipetting all of the primers, master mix, and DNA together, you have to be very careful to not contaminate any of the capsules and to not mix the DNAs together. It can be tricky to remember what you put where with so many different tiny tubes!11124892_737365036372928_2087562588_n

11269517_737379936371438_956346387_nAfter all of the solutions were combined, the capsules were centrifuged and then put into the PCR machine. This machine changes the temperature for specific amounts of time so that the primers and master mix can work to replicate certain sections of the DNA over and over. 11667022_737364883039610_945274925_nAfter the PCR machine was done, we created a gel using agarose. We put combs into the gel to make tiny wells. When the gel had firmed up, it was placed into the electrophoresis machine. 11650707_737364733039625_1993304658_nEach DNA solution had to be carefully transferred into the wells of the gel using a pipette.11651078_737364706372961_599055866_nThe voltage of the electrophoresis machine was turned to 120 V. DNA has a negative charge so it travels towards the positively charged end of the machine. 11647195_737364653039633_1838097742_nThe agarose gel created a matrix for the DNA to move through which made the smaller pieces move farther than the larger pieces.

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Faint yellow and blue lines showed where the DNA traveled

The sheet of agarose was carefully placed into a GelDoc-It Imager.11647335_737364559706309_1299795515_nThe window of the imager allows you to see the gel under UV light.
11668169_737364556372976_1120440093_nThe imager also allows you to record a computer image of the picture to examine for later use. 11124881_737364546372977_997457231_n

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