2. Steps for Southern Blotting
Digest the DNA with an appropriate restriction enzyme.
Run the digest on an agarose gel.
Denature the DNA, which would separate double-stranded DNA into single-stranded DNA.
Transfer the denatured DNA to the membrane.
Probe the membrane.
Visualize your radioactively labeled target sequence. If you used a radiolabeled 32P probe, then you would visualize by autoradiograph.
3. a) She placed the nucleotide as she did because the EcoR I cuts in one place which is why there are two bands. One fragment is slightly smaller than the other.
b) She was able to add this sequence because The Hind III by itself only cut once. The EcoR I also cut once. In the double digest, the Hind III cuts the smaller of the EcoR I fragments.
4. The odds that the sequence will occur in random order is 1 out of 4096. Since there are four bases possible and they must appear in a six base sequence the answer is 1 out of 4.
5. By digesting the......
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