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#1
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| I'm considering doing a lambda plaque hybridization with a very small probe. The unique sequence I'm looking for is 43bp long. I'm lucky to have that much really. We do our colony hybridizations exactly like we would do a southern. So has anyone done a hybridization (plaque or southern) with a tiny probe like this? I imagine it might be hard to get a high enough specific activity, but I think it's worth a shot. |
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#2
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| Hi Ed, when your probe is half size compared to a reference probe then you'll need to double (at least) the specific activity of the probe in order to get the same signal, assuming the probe binds in the same way. This is achieved by increasing the specific activity in the labeling reaction. As you normally will dilute the radioactive molecule with cold compund, just.dilute less. The bigger problem might be to obtain the right stringency for a small probe, so you'll need some fiddling with conditions maybe. HTH Wo On Sep 26, 5:06 pm, Ed Siefker <[Only registered users see links. ]> wrote: |
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| minimum , probe , radioactive , size |
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