1. What is the purpose of pre-hybridization? How would the blotting membrane look like if this...

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1. What is the purpose of pre-hybridization? How would the blotting membrane look like if this...

1. What is the purpose of pre-hybridization? How would the blotting membrane look like if this step was skipped?

2. What is the purpose of blocking? Compare how pre-hybridization and blocking are done.

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Answer 1

Answer #1

1. Pre-hybridization is a blocking step performed before addition of the probe. Its purpose is to minimize non-specific attachment of probe to the membrane.

If this step was skipped, the probe will be binding anywhere on the blotting membrane.

2. Purpose of blocking is to prevent non-specific DNA interactions and reduce background noise.

Washing of the nylon membrane is done with a pre-hybridization solution or blocking reagent comprised of salmon sperm DNA in order to block non-specific DNA interactions. Alternatively, commercial blocking buffers are also available.

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Answer 2

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