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7 (June 7, 2011 GDS)
From 2011.igem.org
(Difference between revisions)
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-Spin 1.5 mL of overnight culture in microfuge </br> | -Spin 1.5 mL of overnight culture in microfuge </br> | ||
| - | -Aspirate all but 100uL of the supernatant and resuspend pellet by vortexing</br> | + | -Aspirate all but 100uL of the supernatant and resuspend pellet by vortexing </br> |
| - | -Add 300uL of TENS and mix by inversion</br> | + | -Add 300uL of TENS and mix by inversion </br> |
| - | -Add 150uL of sodium acetate and vortex</br> | + | -Add 150uL of sodium acetate and vortex </br> |
-Centrifuge for 2.5 minutes at 10K</br> | -Centrifuge for 2.5 minutes at 10K</br> | ||
| - | -Transfer supernatant to clean tube and add 1mL of room temp. ETOH</br> | + | -Transfer supernatant to clean tube and add 1mL of room temp. ETOH </br> |
| - | -Mix and pellet DNA by centrifuge for 2-5 min. at 10K</br> | + | -Mix and pellet DNA by centrifuge for 2-5 min. at 10K </br> |
| - | -Wash pellet with 70% ethanol and allow pellet to dry</br> | + | -Wash pellet with 70% ethanol and allow pellet to dry </br> |
| - | -Resuspend pellet in 30uL of TE w/ RNA seA</br> | + | -Resuspend pellet in 30uL of TE w/ RNA seA </br> |
| - | -Digest 5-10uL as usual</br> | + | -Digest 5-10uL as usual </br> |
Revision as of 19:42, 5 September 2011
-Spin 1.5 mL of overnight culture in microfuge </br> -Aspirate all but 100uL of the supernatant and resuspend pellet by vortexing </br> -Add 300uL of TENS and mix by inversion </br> -Add 150uL of sodium acetate and vortex </br> -Centrifuge for 2.5 minutes at 10K</br> -Transfer supernatant to clean tube and add 1mL of room temp. ETOH </br> -Mix and pellet DNA by centrifuge for 2-5 min. at 10K </br> -Wash pellet with 70% ethanol and allow pellet to dry </br> -Resuspend pellet in 30uL of TE w/ RNA seA </br> -Digest 5-10uL as usual </br>
- To make TENS, add:
- -4.5mL of TE
- -250uL 10%SDS
- -250uL 2N NaOH
- -4.5mL of TE
