α4-200™ plasmid purification kit
$65.00
Catalog: AUK1.
Equivalent of 200 mini-preps.
Our unique kit is designed for purification of up to 58 k.b. plasmid DNA from E. coli cells.
It utilizes a different approach than other commercial kits do.
The entire process of plasmid purification by this kit happens in a single tube and involves fewer steps than micro-column-utilizing kits. Consequently, our kit assures lower per-sample cost of purified DNA and demands less operator involvement time while producing DNA suitable for all types of enzymatic treatments and efficient transformation of all types of cells, including mammalian. Additionally, the kit allows easy up-scaling of the procedure depending on the amount of DNA required.
a4-200™ plasmid purification kit.
Protocol
Prior to using the kit for the first time, add 8 mL of 96% ethanol to the bottle labeled “Solution #3”.
Store the kit in the refrigerator.
1. Collect cells from 1.5 mL of overnight E. coli cell culture by centrifugation in micro-centrifuge (13 rsf) for 2 minutes and remove supernatant (by opening and inverting the tube, then giving it 1 – 2 intense shakes down into a waste receptacle).
2. Re-suspend cells in the remaining (~30 µl) supernatant using a Vortex.
3. Transfer tubes to 80oC and continue incubation for an additional 15 minutes.
4. Add 100 µl of Solution #1 per tube.
5. Thoroughly mix the contents of tubes (short burst on Vortex should suffice) and place the tubes at 37oC for 15 minutes.
6. Centrifuge tubes for 10 minutes at 13 rsf. Some cultures may require a longer spin to achieve a pellet volume of ~ 30-50 µl.
7. Remove pellets from tubes by inserting a sterile toothpick (provided in the kit) into each pellet, spinning it inside of pellet, and pulling the toothpick out of the tube together with the stuck pellet. Note: We recommend using metal forceps sterilized by flame for taking toothpicks out of the storage container.
8. Add 150 µl of Solution #2 to the remaining supernatant and mix the contents by gently inverting the tubes several times (for plasmids larger than 20 k.b.) or by intensely shaking 2 – 3 times.
9. Centrifuge tubes for 10 minutes at 13 rsf.
10. Remove supernatant by aspiration.
Note: We recommend using faucet aspirator/vacuum pump connected to 10 ml pipet tip. Move the tip along the side that is opposite to that where the pellet should be located. We found that the same tip can be used for multiple samples if briefly washed with fresh deionized water.
11. Add 50 µl of Solution #3 (remember to add ethanol) per tube.
12. Centrifuge tubes for 2 minutes at 13 rsf.
13. Remove supernatant by aspiration.
14. Repeat steps 11 – 13 one more time.
15. Add 50 µl of Solution #4.
16. Incubate at 37oC for 5 – 10 minutes.
17. The purified DNA is ready for further manipulations!
AUK1
200 Items
Catalog: AUK1.
Equivalent of 200 mini-preps.
Our unique kit is designed for purification of up to 58 k.b. plasmid DNA from E. coli cells.
It utilizes a different approach than other commercial kits do.
The entire process of plasmid purification by this kit happens in a single tube and involves fewer steps than micro-column-utilizing kits. Consequently, our kit assures lower per-sample cost of purified DNA and demands less operator involvement time while producing DNA suitable for all types of enzymatic treatments and efficient transformation of all types of cells, including mammalian. Additionally, the kit allows easy up-scaling of the procedure depending on the amount of DNA required.