This experiment involved the digestion of Lambda DNA using different combinations of restriction enzymes to analyze fragment distribution through gel electrophoresis. Three lanes were prepared with distinct enzyme mixtures, and the digestion efficiency was recorded. Below is the interpretation of the obtained results.

Lane 1: Digestion with BamHI, EcoRI, EcoRV, HindIII, KpnI, SacI, and SalI

• This lane shows the most fragmented DNA, as multiple enzymes were used simultaneously.
• The highest number of cuts resulted in numerous small fragments, creating a complex banding pattern on the gel.
• Enzyme activities varied:
  • BamHI, EcoRI, HindIII showed moderate to high efficiency (25-100%).
  • EcoRV exhibited lower activity (10-50%), leading to slightly larger fragments.
  • SalI had low (<10%) to full (100%) activity, producing inconsistent cleavage.
  • KpnI and SacI were unavailable, reducing the expected number of cuts.
• Conclusion: The gel pattern is dense, making this digestion ideal for generating intricate Gel Art.

Lane 2: Digestion with EcoRI, KpnI, and SalI

• This lane had fewer fragments compared to Lane 1 because only three enzymes were used.
• EcoRI was highly active (100%), ensuring efficient DNA cleavage.
• SalI exhibited variable efficiency (<10-100%), affecting the consistency of fragment sizes.
• KpnI was unavailable, leading to fewer cuts than expected.
• Conclusion: The larger DNA fragments produced a simpler and more distinct pattern compared to Lane 1.

Lane 3: Digestion with EcoRV and HindIII

• This lane presents a moderate number of fragments, as only two enzymes were used.
• EcoRV exhibited variable activity (10-100%), creating a mix of small and medium fragments.
• HindIII showed moderate efficiency (25-100%), contributing to additional cuts.
• Conclusion: The resulting banding pattern differs significantly from Lanes 1 and 2, demonstrating the unique fragment distribution created by different enzyme combinations.

Key Observations