Haynes Lab:Notebook/CRISPR Editing/2016/09/03: Difference between revisions
Rene M Davis (talk | contribs) (Autocreate 2016/09/03 Entry for Haynes_Lab:Notebook/CRISPR_Editing) |
Rene M Davis (talk | contribs) |
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==03/ | ==09/03/2016== | ||
ChIP incubation ran 12 hours overnight.<br> | |||
Added washed beads to IgG samples<br> | |||
Washed FLAG beads 4x with 1mL TBS buffer, 10 min nutation between washes<br> | |||
after last wash, removed TBS, quick spin, remove remaining TBS with P20<br> | |||
added 100ul elutiion buffer to each sample<br> | |||
Added 50ul elution buffer to inputs<br> | |||
put at 65deg for 8 hours | |||
<br><br> | |||
Washed IgG bead samples after 3 hour nutation at 4deg, 6x with RIPA buffer, 10min nutation between each wash<br> | |||
washed twice with TE pH 7.6, 3 minute nutation between washes<br> | |||
removed TE after last wash, quick spin, removed remaining TE buffer with P20<br> | |||
added 100ul elution buffer to each sample<br> | |||
put at 65deg for 5 hours | |||
<br><br> | |||
30 mins at 37deg with 1ul RNase A<br> | |||
2 hours at 62deg with 0.5ul proteinase K | |||
<br><br> | |||
purify with Sigma columns<br><br><br> | |||
Started some cultures from the g032 plate | |||
:4 1mL cultures for 3 hours | |||
:2mL culture -> 100mL media | |||
:grew for 5 hours | |||
:maxi prep all 100mL | |||
Revision as of 20:18, 3 September 2016
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09/03/2016ChIP incubation ran 12 hours overnight. Started some cultures from the g032 plate
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