Back to timeline

DNA after all?

I ran another mini-prep, although since my centrifuge is funky I used a syringe filter after the precipitation step. Then to see if there is DNA there, I added 2uL SeeGreen stain to 20ml water. I add 200uL of this plus 2uL of my samples: known DNA plasmid at 10ng/uL, miniprep from ag, miniprep from ecoli last time, water (W) and the latest product, this time from liquid culture, e.c. with pBLUE. Did this first in micro centrifuge tubes and couldn't see anything past their own blue scatter/fluorescence. Transferred to glass vials and bingo, finally, a difference between the water and the others. I need to take some better pics, but it looks like they all contain DNA after all! And the color makes me think my fears when I ran the gel and saw the orange clouds were founded: the DNA prob just washed out of the wells. TIL add buffer first! As I was pouring the buffer I noticed surface tension or whatever meant well 3 was the last to fill, at which point I stopped pouring, prob why it was the only one that showed bands. Other likely contributing factor is my wimpy centrifuge not getting rid of as much residual ethanol from the DNA wash buffer during the miniprep, which can make it floaty. Anyway, at least I think I have DNA aplenty now to test electroporation and such with! Maybe in the future I'll build something nanodrop-like to quantify conc. or find a lab with one.