4 replies to this topic

[Curtis]

Hi all,

my boss sent me some plasmids in an envelope and apparently he has just put one drop on a piece of paper for each.

As long as I don't want him to know I'm "unprofessional" and always want to show myself good to him, I don't want to ask him how the heck I'm supposed to elute these plasmids off that paper  

So I'm asking you guys  

Edited by Curtis, 29 June 2010 - 07:06 PM.

[perneseblue]

The process is highly technical and requires great skill that takes years to master. I believe for the average person it takes between 4 to 6 years. Girls (as usual) are able to master this skill faster and at a younger age.  

First, one must acquire a scissors (Always have a responsible adult to supervise) <lol: >.

Usually there will be a circle around the spot where the DNA was placed. Use the scissors to cut a small circle around said spot of DNA. (all those hours, so long ago, learning how to cut paper into funny shapes is finally paying off)

Drop this paper cut out into a micro tube. You can use tweezers to help.

Once in, add about 50ul of TE or water to the paper cut out. You can use a pipette tip to squeeze the paper.

Next place the tube into a centrifuge and spin down for a 30sec or so to pellet the mass of paper.

Take between 0.5-1.0 ul and transform into cells. You will only need to plate a small portion of transformed cells, about 10ul of 1ml. Competent cells are very good at picking up plasmid DNA. When transforming plasmid DNA back into cells, there is a real risk plating to many cells and obtaining a lawn, which can rather quickly degrade any antibiotic on the plate surface.

[Curtis]

my goodness, I feel like I need to go back to elementary school. the whole protocol sounds like a rocket science to me!

[laurequillo]

Just be sure to have at least a biosafety level 2 lab...

[Curtis]

I hope nobody accuses me of making nuclear bomb, I don't want to be on CNN