Griffitts:Miniprep: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
m (→Materials) |
||
Line 4: | Line 4: | ||
<font size="5">Standard Qiagen Miniprep</font> | <font size="5">Standard Qiagen Miniprep</font> | ||
==Materials== | ==Materials== | ||
*Qiaprep Spin Kit (27106) | * Qiaprep Spin Kit (27106) | ||
*TE | * [[Griffitts:Common_buffers#TE_buffer|TE buffer]] | ||
==Procedure== | ==Procedure== | ||
#Grow DH5alpha with appropriate plasmid in liquid LB (4 ml) for 12-18 hours. | #Grow DH5alpha with appropriate plasmid in liquid LB (4 ml) for 12-18 hours. |
Revision as of 12:38, 8 November 2007
Standard Qiagen Miniprep
Materials
- Qiaprep Spin Kit (27106)
- TE buffer
Procedure
- Grow DH5alpha with appropriate plasmid in liquid LB (4 ml) for 12-18 hours.
- Spin down 1.3 ml of culture, dump and tap out supernatant, and repeat.
- Resuspend pellet in 250 ul P1 (from fridge).
- Add 250 ul P2, immediately invert 3 times, let stand for 2 min.
- Add 350 ul N3, immediately invert 8 times.
- Spin 6 min.
- Carefully move supernatant (~820 ul) to spin column.
- Spin 30 s.
- Remove flowthrough and add 650 ul of wash buffer PE to column.
- Spin 30 s.
- Remove flowthrough.
- Spin 60 s.
- Move column to 1.5 ml tube, add 50-100 ul TE directly to disk, let stand 1-2 min.
- Spin 60 s to elute DNA.
Adapted from Qiagen handbook