Exactly how you read them depends on how you have them stored in the file, but if they are on separate lines the easiest way is to use ReadAllLines:
string[] paths = File.ReadAllLines(@"D:\Temp\myPaths.txt");
if (paths.Length >= 2)
{
string sourcePath = paths[0];
string destPath = paths[1];
...
}
If you then want to read from the source:
string data = File.ReadAllText(sourcePath);
Or write to the destination:
File.WriteAllText(destPath, data);
There may be other methods that are more appropriate, depending on what / how you want to read / write but that is up to your application.