|Gel electrophoresis set up including the black death stuffed toy that watched to make sure our gel ran smoothly.|
Tuesday, July 21, 2015
Agarose Gel Electrophoresis!
The past two days, we have prepared and ran 2 gels to determine the best digestion for the chromatin. I added various amounts (from 5 to 80 Worthington units/mg chromatin) of micrococcal nuclease to small samples of the chromatin solution. We made sure that the concentration of DNA in the solutions was less than 100ng/ml so that the gel would run without smudging. We mistakenly let the first gel run overnight at a low voltage but it over ran. Today we ran it through lunch until the visible blue line was at about 2.5 cm. We determined from the imaged gel that 15 Worthington units/mg chromatin of the micrococcal nuclease is sufficient. I then digested all of our chromatin and centrifuged it down. Tomorrow we will be doing a column and another gel to see if our nucleosomes are adequate to use for experiments.