Genetics
Blotting
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Get accessUses a cloned probe to detect presence of a nucleic acid fragment and assess whether it is grossly normal
- These techniques cannot detect single base mutations
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Blotting techniques
- Southern blotting (technique applied to DNA):
- DNA undergoes restriction endonuclease digestion
- The fragments are separated by size via electrophoresis
- They are then denatured using a strong base to form single stranded nucleic acid
- Then incubated on filter paper with a radiolabeled probe (ssDNA)
- Unbound probe is then washed off
- X-ray is used to reveal the positions where the probe hybridised
- Northern blotting is the RNA equivalent
- Same technique except RNA cannot be digested with restriction enzymes
- Western blotting is used to identify proteins