Molecular mechanisms for cutting DNA or RNA can help a cell ensure its safety and future, whether it’s for genetic recombination or in the case of bacteria, using restriction enzymes as a defense mechanism, slicing apart the DNA of a foreign invader. These molecular scissors also allow researchers to study smaller pieces of DNA more thoroughly by being able to cut specific pieces of DNA, or understanding how DNA is organized based on cut sites. In this restriction digestion lab, students will explore what restriction enzymes do, perform single and double digests of DNA, predict fragment sizes and compare to fragment size of the actual digested DNA run on an agarose gel, and use pre-digested DNA controls. Appropriate for AP, honors and advanced biology students, grades 9-12.
Enzymes should be stored in the freezer.
GreenGel GelCups should be left in the original package and protected from light; guaranteed stable for six months with proper storage
Materials Included in Each MiniLab:
Each MiniLab contain s enough materials for 10 workstations, 2 – 3 students per workstation.
- Ten 1.5% agarose GreenGel™ Cups
- Undigested DNA Sample
- Three pre-digested DNA controls
- MiniOne® Universal DNA marker
- Enzyme dilution buffer
- Two restriction enzymes
- MiniOne® 5X Sample Loading Dye
- One bottle of 100 mL Tris-Borate-EDTA (TBE) buffer concentrate, enough to make 2L of 1X running buffer
- One bag of 0.65 mL microcentrifuge tubes
- One bag of 2 – 200 µL micropipette tips
- One bag of 0.2 mL PCR tubes
- Teacher’s guide
NOTE: This item contains temperature sensitive reagents that will ship separately. Temperature sensitive reagents need to be stored in the freezer immediately upon receipt.
Equipment required (not included) for this MiniLab:
PCR machine or water bath. See Teacher’s Guide* for quantities and complete list of materials needed for this activity.
Utilize these teachers guides and classroom handouts to get the most out of your restriction digestion MiniLab.