DIY Bacterial CRISPR Refill/Classroom Kit
Posted by Catherine on 12th Dec 2018
Have run this successfully in sophomore biology for the last three years. I wish it was possible to do a little more a la carte when purchasing. I'm in a high school science lab so I don't need everything, but the refill kit didn't have everything I need. I bought 2 kits (should be 10 experiments) for 6 groups but the 3 chemicals that get added to the competent cell mixture ran out after 5 groups.
successfull class runs
Posted by S. Caldwell on 4th Dec 2018
We were able to make this fit into a 3 hour lab successfully with only a 1 hour outgrowth time at 30 degrees c. Many groups had too much bacteria...just 1 colony from the stock plate will do!
Classroom Kit - I'm going to try for 4 stations per kit
Posted by Sara Caldwell on 15th Nov 2018
I have just gotten 2 of these kits for classroom use. I am used to having to get a quote for shipping, but the flat $20 covered both kits and made estimating costs easy. For others who maybe trying to figure out how much to get/what is in each here is the list for one kit: enough dry LB agar for 150 ml , enough dry LB antibiotic agar for 150ml, a 50ml
tube for measuring, 14 petri dishes, 5 disposable loops, 1 disposable cell spreader, 1 stab culture of non-pathogenic E.coli, 6 pre-measured microtubes of dry LB media, ~1ml transformation solution, ~ 20 empty microtubes, 4-5 pairs of gloves, 55 ul each of the 3 Crisper ingredients. Though one person using a kit might get 5 runs, I'm going to split this into four stations going at once. I also noticed that each run makes enough inoculate for not just one but three plates. I practiced with nutrient agar and discovered if I pour thin and swirl can get 8 plates out of 150 ml ...ie enough for each group to prepare 2 plates. The second set of plates will come in very handy because I want to test if we can trim time from somewhere in this protocol. I will need to fill in with more gloves, crushed ice, beakers of disinfectant, pipettors, waterbath and some more cell spreaders but overall I think this will work well. I will review again after the lab.
Nobel Prize work in a classroom!
Posted by David R on 23rd Oct 2018
Wow! I've done this successfully for 2 years now. I love sharing with my kids that we can MODIFY THE DNA of an organism. It makes the dots you see on an agar plate have so much meaning. Thanks for offering this!
Great for the Classroom
Posted by George on 11th Jul 2017
This kit was a success, directions were easy to understand and would definitely purchase again!
Hard to understand
Posted by Gage on 4th Feb 2017
This was a really go experiment to do. It was hard to understand the instructions though and the only way I knew what to do was to talk to my professors. On top of all that we were all afraid that the E. coli was pathogenic and was going to kill us all. We did not see that it was non pathogenic until after the experiment. Other than all that though it was a thoroughly engaging, interesting, and successful experiment. It helped me to figure out that I want to get a microbiology degree.
This kit provides the media, DNA, bacteria and plasticware to perform a bacterial CRISPR experiment. 1 item is enough for 5 experiments.
- 50uL of Cas9 plasmid
- 50uL of gRNA plasmid
- 50uL template DNA
- Strep/Kan/Arabinose Agar Media
- LB Agar Media
- LB Media
- 20 microcentrifuge tubes
- 14 plates
- Non-pathogenic DH5a E. coli bacteria
- Inoculation loops
- 750uL Bacterial Transformation Buffer
*This kit does not contain pipette, pipette tips and glass bottle for making media. These must be user supplied.
Some items in this kit need to be stored in a fridge and a freezer upon you receiving them.