Southern Blot/ How do you clean hybridization tubes

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suntoucher
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Southern Blot/ How do you clean hybridization tubes

Post by suntoucher » Wed Jan 18, 2006 5:35 pm

I have never used southern before. People showed me how to hybridize membrane with 32P labeled probe in the sealed bag but it looks VERY MESSY. Especially when you're trying to remove bubbles from hybridization solution which is in the bag and radioactive probe is already there so that is going outside on your gloves... It's so inconvenient and unsafety even it's has been done in the hood...

Likely we got Hybridization Incubator and it has rottator or "rottisier" with 2 kind of tubes for rotation. There's my q-n: how do you do "rotation method" for hybridization? Do you put membrane and hybridization solution w/32P-labeled probe inside that original tube? How do you clean a tube after that?

I tried to put Corning 50 ml tube into the tube from Incubator but it doesn't fit. :roll:

Thanks in advance.

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canalon
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Re: Southern Blot/ How do you clean hybridization tubes

Post by canalon » Wed Jan 18, 2006 8:16 pm

suntoucher wrote:I
Likely we got Hybridization Incubator and it has rottator or "rottisier" with 2 kind of tubes for rotation. There's my q-n: how do you do "rotation method" for hybridization? Do you put membrane and hybridization solution w/32P-labeled probe inside that original tube? How do you clean a tube after that?


You need to use the special tubes because all others may become leaky after too long at high temperature. But yes you basically just carefully put your membrane in the tube (roll it and let the bottom side stick to the walls, if it is too big and there is some overlaping, just but some nylon clothe under the membrane before rolling) and add the pre hybridization solution, set your rotisserie at the correct temp, and after a while replace it with the hybridization solution. Et voila.
But I don't know how to safely wash the tube. That is why I definitely prefer non radioactive systems like DIG... And that is probably why even though messy bags are used: they just go to the radioactive waste. And if done properly, it is usually not that messy (when I use bags I prepare a channel for emptying?filling them with the heating thing....
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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