Problems with DIG DNA labeling! help! - Membrane was completely stained purple with no bands (Sep/23/2007 )
QUOTE (Lene @ Oct 2 2007, 08:57 AM)
Hi all...
I tried to reuse the Hyb soln and the good news is that it still worked. I didnt cool it down after heating it but straightaway poured it onto the membrane however I think that it would produce better yield if it was cooled down after heating before use. I was wondering whether the prehyb can be used? so far i've not heard anyone that reuses their prehyb, so i was wondering if anyone has ever tried that before.
Cheers,
Charlene
I tried to reuse the Hyb soln and the good news is that it still worked. I didnt cool it down after heating it but straightaway poured it onto the membrane however I think that it would produce better yield if it was cooled down after heating before use. I was wondering whether the prehyb can be used? so far i've not heard anyone that reuses their prehyb, so i was wondering if anyone has ever tried that before.
Cheers,
Charlene
Alright, Thanks for the information. I think I will probably try it out too...for my hybridization. Well, I just accidentally throw away my hyb solution 
. Anyway, I still kept some other hyb solution. Thanks once again.I oso wondering will the pre-hyb and also the antibody solution can be reuse? From the topic I posted, Phage told that Anti-Dig can be reuse, but is not advisable to do so. I wonder 
... Thanks. -cheerioet83-
This is why you have controls. If the controls are working with the reused solutions, then things are fine, and you can continue to work with them. If the controls don't work, then you need to find the problem. This could be one of the potential problems. But until your controls are working, you are foolish to be working with anything (additional) that might make things not work.
-phage434-
QUOTE (phage434 @ Oct 3 2007, 08:41 AM)
This is why you have controls. If the controls are working with the reused solutions, then things are fine, and you can continue to work with them. If the controls don't work, then you need to find the problem. This could be one of the potential problems. But until your controls are working, you are foolish to be working with anything (additional) that might make things not work.
Uh.ya..Guess that you are right. After seeing my control working, I do enlighten me and prove to me that it isn't my technique or my protocol fault. But somehow, it is really headache when my control didn't work, especially for my other probe..where the labeling efficiency test shown a good result, but it ain't bind to my reference sample. Which means, the probe isn't working for my sample, right? 
Well, hybridization seems to be my work for the coming weeks. Sigh...
-cheerioet83-