Southern blot membrane cross-link
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Southern blot membrane cross-link
A friend of mine wants me to find out if nylon membranes can be baked without
a vacuum to cross-link DNA onto the membranes.
WE do it all the time! Seems to work in a regular oven just fine.
This has been said before (sorry, don't remember who) but I'll say it this
time. The _only_ reason vacuum ovens were used in the past is to keep
nitrocellulose paper from bursting into flame (heat + oxygen + nitrocellulose
= ash ). Any non-flammable paper/membrane, such as nylon, is OK to bake
without vacuum. (Having said that, in my lab we use the vacuum anyway. Go
figure. :-) )
Jim
Baking will immobilize DNA on a nylon membrane. You dont need a vacuum oven
because nylon membranes do not have a high flash point the same way that
nitrocellulose (AKA guncotton) does. The baking will hold the target DNA just
about as well as UV crosslinking. Biotechniques had an article about this a
while back. I think the lead author was Sandra A. Nierzwicki-Bauer.
Tell your friend to go for it
Last update 16-Aug-2001, Rating Fair of 1 votes.
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I am getting problem in southern hybridization. I got the same signal pattern in different transgenic lines, the genomic DNA was cut with one enzyme in all.What can be the reasons?
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what should be the ideal concentration prode dna
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hkjhgkjh
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I prefer UV linking to fix the DNA on the membrane. It takes all of 10 seconds and is very easy to set up. We've had good results using this method.
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