dna hybridization

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Gozza
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dna hybridization

Post by Gozza » Mon Sep 05, 2005 8:57 pm

Hi there. I need to do some hybridization of complementary oligos but I have some (probably stupid :oops: ) questions.
1. Do I need to use these nitrocellulose membranes? My idea was to just to mix two complementary strands.
2. Is there any oligo concentration dependence of hybridization?
3. How do I monitor when that hybridization occurred (i.e. is there any “real time” signal (maybe like absorbance) that I can monitor to see when the hybridization occurred.
4. Would the hybrid DNA (it’s around 20 bp) survive sonication?

Any other advice to poor physicist making his first steps in biology is welcomed :P

weesper
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Post by weesper » Tue Sep 06, 2005 7:43 am

You would have to indicate what kinf of exp your looking for; the nitrocellulose sounds like you're doing a Southern but the 20bp sounds more like you're preparing oligos for cloning and doing an RNAi exp; if the latter is the case, just hybridizein waterbath; turn off waterbath (slow cooling helps the oligos anneal nicely) and then clone that into your construct; no need checking for hybridization just digest the construct after ligating and trace the products on a gel. If this is not what you're looking for let me know. gr weesper

Gozza
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Post by Gozza » Tue Sep 06, 2005 5:37 pm

Thanks a lot weesper :D . I am doing just simple hybridization so I guess this hybridization waterbath will work.

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