Ran ~250ng (out of 3000ng, according to Claire) of LSU C.virginica oil spill gDNA on a gel that was previously sheared by Claire to verify that shearing was successful.

Ran unsheared side-by-side with sheared gDNA for comparison.

Note: HB16 and NB3 did not have any unsheared gDNA left in their tubes, so nothing was run on a gel.

Results:

Ladder used: O’GeneRuler 100bp Ladder (ThermoFisher)

Well, it’s rather obvious that the initial shearing did NOT work. Will re-shear the samples.

UPDATE: Looking at the Biorupter (Diagenode) manual, it turns out that shearing samples in a 1.5mL tube (in which these were sheared) requires a minimum volume of 100uL. All the samples were far below this minimum volume. Additionally, the recommendations in the manual to reach the target size range are significantly longer (30 – 40 cycles) than what was applied (4 cycles). The combination of these two factors are likely the reason that shearing didn’t take place.