pClone: Exploring Promoters with Synthetic Biology Kit (with perishables)

Advanced—For experienced high school and college classes; requires some technical skill.
 
Unique introduction to synthetic biology
Hands-on exploration of the role of promoters in gene regulation
Seamlessly blends together multiple areas of biology
Takes a traditional transformation lab to the next level
Lesson can be facilitated in 6 lab sessions, at most
 
Bring the unique area of synthetic biology into your classroom with this one-of-a-kind kit activity. Your students get the opportunity to take on the role of genetic engineers as they clone promoters into 2 different plasmids. This hands-on approach to transcription regulation exposes your class to a wide array of biological disciplines including biotechnology, molecular biology, and evolutionary biology.
 
Students clone the promoters into the plasmids, pClone Red and pClone Blue, using a common cloning technique, Golden Gate Assembly (GGA). After cloning in the promoters, students are tasked with performing a transformation on E. coli cells to produce new ampicillin-resistant colonies expressing their designed plasmids. Based on the orientation of the promoter, students then observe colonies that appear green (produce GFP), red (produce RFP), or blue (produce AmilCP blue). This gives them 3 distinct colonies to further assess using ImageJ software, freely available, to determine the level of protein expression.
 
The kit includes enough materials for 8 groups of 2 to 4 students. The activities typically take 3 to 6 class periods for completion. Note: Kit comes with the perishable materials included.
 
FREE 1-year access to digital resources that support NGSS instruction
included.

What’s Included:
4 Ampicillin, 10 mg/mL 
1 BsaI 
1 E. coli with Plasmid Containing Plac Promoter and RFP Gene 
1 E. coli with Plasmid Containing P5 Promoter and AmilCP Blue Gene
1 T4 DNA Ligase 
1 10x Ligase Buffer 
1 MM294 E. coli Slant 
1 Mutant Oligonucleotide Pair (P5-33A top and bottom) 
1 Wild-Type Oligonucleotide Pair (P5 top and bottom) 
1 pClone Red Plasmid, 25 µg/µL 
1 pClone Blue Plasmid, 25 µg/µL 
1 10x Oligonucleotide Annealing Buffer 
8 Calcium Chloride, 50 mM, 3 mL
30 Disposable Spreaders, Sterile
30 Disposable Inoculating Loops, Sterile
4 LB Agar, Sterile, 400 mL
8 LB Media, 3 mL
60 Petri Dishes
25 Transformation Tubes, 15 mL
1 Teacher’s Manual with Reproducible Student Guide
 
Needed But Not Included:
 Aluminum Foil
 Beaker, 1 L
 Computer
 Digital Camera
 Disinfectant (ethanol or household bleach solution)
 Hot Plate or Bunsen Burner
 Ice
 ImageJ Software (free download)
 Incubator, 37° C (optional)
 Lab Marker
 Microcentrifuge
 Microcentrifuge Tubes, 0.5 mL
 Micropipettors
 Micropipette Tips, Sterile
 Parafilm®
 PCR Tubes, 0.5 or 2.0 mL
 Programmable Thermal Cycler
 Thermometer
 Tongs
 Tube Racks, for 0.5-mL Tubes
 Tube Racks, for 15-mL Tubes
 UV Transilluminator
 White-Light Transilluminator (optional)
 Water
 Water Bath, 42° C
 Wire Inoculating Loop
 
Shipping Information or Purchase Restrictions
This item contains living or perishable material and ships via 2nd Day or Overnight delivery.