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Question

In restriction mapping, how do you distinguish tandem insertions?

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Explanation

Tandem insertion duplicates a segment in same orientation directly adjacent to original, increasing distance between flanking restriction sites by size of duplication. Upon digestion with enzymes flanking region, fragment containing insertion migrates slower, showing higher molecular weight compared to wild-type. Orientation does not change size but may affect internal sites. Number of enzymes, western blot, gel drying are unrelated. Thus observing band migration shift upward indicates larger insert while retained restriction pattern confirms tandem nature rather than inverted or random insertion.

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