7 (June 7, 2011 GDS)

(Difference between revisions)

Revision as of 19:43, 5 September 2011

-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 sodium acetate and vortex

-Centrifuge for 2.5 minutes at 10K</br>

-Transfer supernatant to clean tube and add 1mL of room temp. ETOH

-Mix and pellet DNA by centrifuge for 2-5 min. at 10K

-Wash pellet with 70% ethanol and allow pellet to dry

-Resuspend pellet in 30uL of TE w/ RNA seA

-Digest 5-10uL as usual

To make TENS, add:

-4.5mL of TE

-250uL 10%SDS

-250uL 2N NaOH

@@ Line 1: / Line 1: @@
--Spin 1.5 mL of overnight culture in microfuge </br>
+-Spin 1.5 mL of overnight culture in microfuge
--Aspirate all but 100uL of the supernatant and resuspend pellet by vortexing </br>
--Add 300uL of TENS and mix by inversion </br>
+-Aspirate all but 100uL of the supernatant and resuspend pellet by vortexing
--Add 150uL of sodium acetate and vortex </br>
+-Add 300uL of TENS and mix by inversion
+-Add 150uL of sodium acetate and vortex
 -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>
+-Transfer supernatant to clean tube and add 1mL of room temp. ETOH
--Wash pellet with 70% ethanol and allow pellet to dry </br>
--Resuspend pellet in 30uL of TE w/ RNA seA </br>
+-Mix and pellet DNA by centrifuge for 2-5 min. at 10K
--Digest 5-10uL as usual </br>
+-Wash pellet with 70% ethanol and allow pellet to dry
+-Resuspend pellet in 30uL of TE w/ RNA seA
+-Digest 5-10uL as usual