Jonesma Lab:Protocols:miniprep: Difference between revisions
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(New page: #Take an overnight <i>E. coli</i> culture and spin down at 3000 rpm #Discard supernatant and resuspend pellet in 250μL P1 buffer #Add 250μL P2 buffer and mix #Incubate at room temperatur...) |
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#Take an overnight <i>E. coli</i> culture and spin down at 3000 rpm | #Take an overnight <i>E. coli</i> culture and spin down at 3000 rpm for 15min | ||
#Discard supernatant and resuspend pellet in 250μL P1 buffer | #Discard supernatant and resuspend pellet in 250μL P1 buffer | ||
#Add 250μL P2 buffer and mix | #Add 250μL P2 buffer and mix | ||
Latest revision as of 01:35, 25 November 2015
- Take an overnight E. coli culture and spin down at 3000 rpm for 15min
- Discard supernatant and resuspend pellet in 250μL P1 buffer
- Add 250μL P2 buffer and mix
- Incubate at room temperature for 4min
- Add 350μL N3 buffer
- Spin at 13 000rpm for 15min
- Take supernatant and add to blue miniprep column (with clear plastic reservoir attached)- maximum volume 700μL
- Spin at 13 000rpm for 1min
- Empty reservoir and add remainder of supernatant from step 7
- Spin at 13 000rpm for 1min
- Add 700μL PE buffer and incubate for 1min
- Spin at 13 000rpm for 1min
- Empty reservoir
- Spin at 13 000rpm for 1min
- Transfer blue column to new 1.5mL tube, labelled appropriately
- Add 40μL dH2O and incubate at room temperature for 1min
- Spin at 13 000rpm for 1min
- Keep tube with DNA, discard used blue column
