Hello to everyone!
For the experts of the DNA fiber technique, I am having issues with my fibers. As you can see in the picture, the fibers are all together as a mass and they have not separated nicely. I have had beuatiful fibers in the past but now I am unable to reproduce them (I have not changed the technique or any single reagent whatsoever!). I am aware that the envrironment does play a role for the experiment so I have tried to conduct the experiment in a warm ish place. It looks like either a) there are too many fibers (they should be 8x105 as many protocols state) or b) the lysis buffer has not worked? I am using the technique from this paper. https://www.sciencedirect.com/science/article/abs/pii/S0076687917301143
The cells that I am using is HeLa, a widely used cell line for this tecnhique. I also make sure that the fibers ''travel'' slow enough (5 mins) along the slide because some papers mention that when the fibers go down quickly, they dont get separated therefore they are not analysable (the problem I have!)
Any help would be much appreciated!
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