7 (June 7, 2011 GDS)

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Revision as of 19:42, 5 September 2011 by Sallen (Talk | contribs)

-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