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