Practice question

Restriction maps order recognition sites along DNA by analyzing fragment overlaps from single and double digestions on agarose gels. If enzyme E1 lies physically between E3 and E4, digestion with E3 and E4 would generate fragment where E1 site resides entirely inside that interval, and triple digest would split that interval into two smaller pieces whose sizes sum to original. Correct map representation showing relative order helps predict fragment sizes for diagnostic digests and cloning strategies. Precise ordering is critical for designing double digests, assembling contigs, and interpreting RFLP polymorphisms in genetic analysis.