Hi all, Thanks for sharing your thoughts. I hope to study the overhangs of the DNA; therefore, need to keep DNA in native conditions. Tracy On Thu, Jul 17, 2008 at 1:25 PM, Tracy Chow <ttchow@gmail.com> wrote: ---------- Forwarded message ---------- From: Jose de las Heras <josenet@tiscali.co.uk> Date: Thu, Jul 17, 2008 at 4:14 AM Subject: Re: DNA crosslinking on nylon

"TC" <tracyscience@gmail.com> wrote in message news:mailman.777.1216254116.3533.methods@net.bio.net... Hello, I am trying to crosslink DNA to nylon membrane for southern blots. Besides traditional UV crosslinking (may induce DNA damage) and baking (causes DNA to denature), are there any other way to crosslink DNA onto nylon memberanes? For my purpose, I hope minimize DNA

Hi Tracy, I think DNA binds pretty strongly to positively charged membranes. I seem to recall several occasions where, using charged membranes, crosslinking was omitted but didn't seem to make any discernable difference... HTH, - Rory -----Original Message----- From: methods-bounces@oat.bio.indiana.edu [mailto:methods-bounces@oat.bio.indiana.edu] On Behalf Of TC Sent: 17

Tracy Chow wrote: Hi Rory, Thanks for your suggestion...really appreciate. May I ask if you use Hybond-N+ or Hybond-XL (also positively charged) membrane? May I ask for the conditions you dry the membrane (room temperature or vacuum)? How long do you *dry the membranes *prior to hybridization? How long do you *pre-hybridize* prior to probe incubation? THANKS!!!