can one over-crosslink nucleic acids to a membrane

colossus... s535290 at
Mon Oct 6 19:30:27 EST 1997

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 ( 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 ?



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