I am attempting to elute a 9kb plasmid from a square of filter paper sent to me to be transformed into DH5a E. coli (photo below). Each filter paper square has been spotted with 1ug of plasmid DNA.
I have two questions:
AddGene's protocol states to elute the DNA using 30ul of TE (I'm using H20 as that's all I have atm). Using a similarly sized piece of filter paper to test, I added it + 30ul of h20 to a 1.5ml tube with a quick 4k spin down and have observed that the 30ul is completely absorbed by the paper as shown in the photo. How should I resolve this? Add an additional 30ul? I also tried to use a pipette tip to shove the paper down and squeeze out the water but you can see it didn't work.
What volume should I use for transformation knowing that the filter paper contains 1ug and I need to use additional elution volume? Most protocols I've read call for 1-5ul of DNA and AddGene calls for 2ul assuming 30ul elution volume.