Team:Gaston Day School/7 July 2009

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(New page: <html> <du> <dt>Spin 1.5mL of overnight culture for 30 sec (5') in microfuge</dt> <dt>Aspriate oft all bu 100h of the supernatant and resuspend the pellet by vortexing</dt> <dt>Add 30x of ...)
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<du>
<dt>Spin 1.5mL of overnight culture for 30 sec (5') in microfuge</dt>
<dt>Spin 1.5mL of overnight culture for 30 sec (5') in microfuge</dt>
-
<dt>Aspriate oft all bu 100h of the supernatant and resuspend the pellet by vortexing</dt>
+
<dt>Aspriate off all but 100h of the supernatant and resuspend the pellet by vortexing</dt>
<dt>Add 30x of TENS and mix by inversion. The solution should become viscous.</dt>
<dt>Add 30x of TENS and mix by inversion. The solution should become viscous.</dt>
<dt>Add 150x of sodium acetate and vortex. A fine, white precipitate should form</dt>
<dt>Add 150x of sodium acetate and vortex. A fine, white precipitate should form</dt>

Revision as of 14:12, 20 July 2009

Spin 1.5mL of overnight culture for 30 sec (5') in microfuge
Aspriate off all but 100h of the supernatant and resuspend the pellet by vortexing
Add 30x of TENS and mix by inversion. The solution should become viscous.
Add 150x of sodium acetate and vortex. A fine, white precipitate should form
Centrifuge for 2.5 min at 10k
TRANSFER the supernatant to a clean tube and add 2 volumes (1mL) of room temperature EtOH
Vortex and pellet DNA by centrifugation for 2-5 minutes at 10k
Wash pellet with 70% ethanol and allow the pellet to dry
Resuspend the pellet in 30x of TE with RNAseA
Digest:
10x DNA
3x Buffer
15x H2O
2x Xba
2x pst
Threw out RFP
Made more TENS

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