Question

in regards to lane 3 of this gel, which band is nicked, linear and supercoiled?

Lysis & Boiling Method Preps (Students) 45 ked ectrophoresis ests) wolts a nicked ) linear supercoiled 1% Agarose Gel Graduate Students Alkaline Lysis-Boiling Method Preps EcoRI-BamH-I Digests 02-14-2020 A2188

Answer 1

In lane 3, top most band from loading point (sample loading side) is linear as after digestion with restriction enzyme it's become linear so occupied more surface area and hence slow running.

Below than linear band, is band of nick one in which it's one strand is relaxed while other is supercoiled so occiped average surface area due to partial digestion of plasmid due to restriction enzyme.

While lower most band is of supercoiled as plasmid is still circular and hence surface area is also less so running fast.

Hope it's clear .thanks