I realized a plasmid extraction with miniprep kit (FERMENTAS).
Yield: 44,9ng/uL by qubit 2.0
So I realized a restriction assay with Hind III for linearization of my standard. I followed the manufacture's recomendations of the enzyme.
I used 10uL of plasmid sample [10 *44,9], with final volume of 20uL
So for gel purification i tried 8% agarose gel + 7uL of my sample (linearized - " I expected").
Run time: 1:20h
Expected size = 5700 pb (vetor + insert)
But I can't find nothing in my gel.
Questions: Is this concentration [44,9 ng/uL] a problem? Have I to try another extraction for a better yield?