can one over-crosslink nucleic acids to a membrane

cgriffin at wco.com cgriffin at wco.com
Thu Oct 9 18:32:01 EST 1997


Hello Ed,

Just a thought, in theory, excessive exposure to UV for cross-linking can
damage the target, and thereby compromise subsequent hybridization
results. I for one would suggest that 40 min. is far to long period. Why
not try baking at 80 degrees which, if you are using nylon, can be done in
excess...no problem. 

As I said, just a thought

C. Griffin
SFGH: GCRC Mol. Biol. Core




In article <Pine.A41.3.95.971006202347.87434J-100000 at aix1.uottawa.ca>,
"colossus..." <s535290 at aix1.uottawa.ca> wrote:

> Hi all,
> 
> I know that people sometimes calibrate their UV source, which implies that
> either "too little" or "too much" cross-linking would be detrimental to a
> later hybridization. Let's just sake for the sake of argument (yeah...or
> let's say this scenario happened to a "friend" :)) that one should leave a
> blot crosslinking for way too long (umm....by about 40 minutes !!!!), are
> these blots toast ? pardon the pun...they're colony blots, which are
> certainly much more robust than northerns or southerns, so is it still
> worth my while to go ahead and use these ? or am I likely to be wasting my
> time ?  it's just that I picked and patched about 300 colonies for this...
> and I REALLY would rather not have to re-do it unless I have to. But the
> thought of going through all that radioactivity jazz for the sake of blots
> that are not going hybridize is also a distasteful consideration...what
> say ye' folks ?
> 
> TIA,
> 
> Ed



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