What does this look like to you? - (Mar/06/2009 )

Thanks for your help, but i really want to be able to do this with a large amount of DNA, say 3ug. The gel purification doesnt give that good of return on my product, like 50%. I want more since i have to use for quantitation and digestion and i want to make it really concentrated product as well. My question is how do you compromise the volume of the digestion and running the gel to purify it. You get a huge band that requires alot of gel in return requires alot of binding buffer for binding to the column. Do you understand what im saying. Your help has been much appreciated, kept my from pulling my hair out, lets hope the ligation goes well.

I just want to have alot of DNA, and concentrated. I bought these clonables novagen dna ligase master mix which requires 5 ul of plasmid and insert total and 5 ul of master mix. So i need concentrated plasmid for this (50ng/ul or so).