Description
Knockout! A CRISPR/Cas Gene Targeting Lab
This lab is easy for teachers to prepare, makes gene targeting accessible to students, and packs a powerful punch!
Become a genetic engineer and use CRISPR/Cas to disable a gene in bacteria. Scientists use the CRISPR/Cas system to alter the DNA in living cells. Now you can bring this cutting-edge technique to your a
dvanced biology class! In this lab, E. coli cells which express the lacZ gene from a plasmid will be transformed with another plasmid containing the cas9 gene and a custom guide RNA. This guide RNA contains a sequence complementary to the lacZ gene so, in transformed cells, the guide RNA will target Cas9 to cut the lacZ gene, disabling it. Students can easily identify their CRISPR/Cas knockouts using the color of the bacterial colonies. Cells which express lacZ are able to break down the chemical X-gal into a blue product. Cells with a functional lacz will be blue, while cells in which the CRISPR/Cas system has successfully disabled lacZ will revert to a white phenotype.
Note: This is a technically advanced lab that requires a 2-hour class period. Students should have prior experience with bacterial transformation.
- Techniques: Micropipetting, bacterial transformation, gene targeting
- Topics: Genome editing, biotechnology, molecular biology
- Time required: Requires a 2-hour class period
- Level: Advanced high school through college
This lab provides enough reagents for eight lab groups:
- Plasmid DNA
- pCtrl DNA
- pKO DNA
- Calcium chloride
- Lyophilized DH5alpha E. coli with lacZ plasmid
- LB ampicillin
- SOC recovery medium
- ChIX Mix
- LB agar
- 20 sterile Petri dishes
- 25 sterile spreaders
- 100 sterile 1.7 ml tubes
- Knockout! sticker
Required equipment:
- Micropipettes and tips
- Water bath or dry bath
- Incubator
Storage notes:
- Some reagents require refrigerator storage and some require freezer storage
- Reagents are stable for 3 months when stored as directed
Refill kit available:
- Does not include necessary plasticware and LB-agar media,
Knockout! A CRISPR/Cas Gene Targeting Lab (Refill kit)
Refill kit, does not include necessary plasticware and LB-agar media.
This lab is easy for teachers to prepare, makes gene targeting accessible to students, and packs a powerful punch!
Become a genetic engineer and use CRISPR/Cas to disable a gene in bacteria. Scientists use the CRISPR/Cas system to alter the DNA in living cells. Now you can bring this cutting-edge technique to your advanced biology class! In this lab, E. coli cells which express the lacZ gene from a plasmid will be transformed with another plasmid containing the cas9 gene and a custom guide RNA. This guide RNA contains a sequence complementary to the lacZ gene so, in transformed cells, the guide RNA will target Cas9 to cut the lacZ gene, disabling it. Students can easily identify their CRISPR/Cas knockouts using the color of the bacterial colonies. Cells which express lacZ are able to break down the chemical X-gal into a blue product. Cells with a functional lacz will be blue, while cells in which the CRISPR/Cas system has successfully disabled lacZ will revert to a white phenotype.
Note: This is a technically advanced lab that requires a 2-hour class period. Students should have prior experience with bacterial transformation.
- Techniques: Micropipetting, bacterial transformation, gene targeting
- Topics: Genome editing, biotechnology, molecular biology
- Time required: Requires a 2-hour class period
- Level: Advanced high school through college
If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200
