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SEPGuides: Science Education Partnership: DNA Restriction Enzyme Digest

SEPGuides is the go-to place for all SEP kit protocols and activities, as well as the posting area for announcements and program updates. If something is missing or you have an idea to contribute, please e-mail us at sep@fredhutch.org

DNA Lab 2 - Restriction Enzyme Digest

DNA restriction enzyme digest is a useful tool when analyzing DNA. In this lab students will be performing their own enzyme digest and comparing the resulting band patterns in their gels.

DNA Lab 2 Resources

Editable Versions

Protocol Cards

Additional Materials

Enzyme Restriction Resources

Enzyme Restriction Extras


NEB's video on Type II Restriction Enzymes

Extra Resources

Here is a very cool time lapse video of gel electrophoresis in action, courtesy of retired SEP teacher and current RV explorer extraordinaire Aram Langhans.

Aram used the FlashGel System, created by Cambrex, and the actual running time was only 5 minutes. More info atwww.flashgel.com.

Save Time in the Lab

A significant reduction in gel electrophoresis running time can be achieved by decreasing the concentration of the running buffer (TAE) and increasing the voltage:

  • Decrease the running buffer from 1X to 0.25X
  • Increase the voltage from 100V to 200V
  • Reduce your running time from 45 minutes to 20 minutes.

BE SURE YOU:

  • Make the agarose with 1X TAE NOT the 0.25X running buffer
  • Do NOT run the gel longer than 20 minutes.

If you exceed 20 minutes the gel will melt.

Under these conditions, the top edge of the gel (where the wells are located) may begin to melt a bit, but this should not impact the visualization of DNA fragments.

- Tip courtesy of Essy Levy of Bio-Rad