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Answer for 1c)

If we are unable to get proper signal in our southern blot then we should find ways to enhance the sensitivity. To increase the sensitivity means to increase the signal or eventually the number of probes (labels) in specified areas. Hence, we can digest more DNA and then load it per lane. Probe is generally taken in excess and I think taking more probe would not affect the sensitivity, but if we think that we had taken a less concentration of probe, then we should increase it. Narrower lane would eventually means reducing the total volume of DNA that can be loaded. If we think that we are taking DNA that gets “diffused” then we can reduce the lane diameter, else, I do not think it is a crucial parameter. Smaller volume of hybridization would not affect the sensitivity unless there is an enhanced amount of probe used. Hybridization time can be increased that would provide more chance for the probe molecules to bind with their target. However, we should note that the number of molecules of probe bound would only be up to the total number of DNA molecules available on the membrane; hence once saturated, we cannot enhance the signal by increasing the time of incubation....

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