Kafatos:Minipreps
| Click here to visit our NEW WEBSITE |
| The content below is most likely out of date. We also have a new lean and mean openwetware area. |
Day 1
- inoculate 5 ml sterile LB medium (+ antibiotic ?) with single bacterial colony
- grow ON @ 37˚C while shaking vigorously
Day 2
- spin 1.5 ml 2’ @ 8,000 rpm in table top centrifuge
- resuspend pellet in 100 μl P1 Buffer. Let sit 5’ @ RT
- add 200 μl P2 Buffer, mix and let sit no more than 5’ @ RT
- add 150 μl ice cold P3 Buffer, mix well and place 5 – 10’ on ice
- spin 10’ @ max rpm (4˚C if possible)
- transfer supernatant to new tube and add 2 volumes of abs EtOH. Let sit for 10 – 20’ on ice
- spin 10’ @ max. speed @ 4˚C
- discard supernatant and wash pellet with 1ml of 70% EtOH
- spin 5’ @ max speed
- suck up or decant supernatant, speedvac or dry pellet @ 37˚C (do not overdo it, though, or it will not properly dissolve again)
- resuspend pellet in 30 μl TE Buffer or ddH20 and check on gel
note: plasmids obtained with this protocol are not clean enough for sequencing
BioStream version
Following is the Kafatos:Minipreps protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.Vaishnavi
