Haynes Lab:Notebook/CRISPR Editing/2016/09/03

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09/03/2016 ChIP incubation ran 12 hours overnight. Added washed beads to IgG samples Washed FLAG beads 4x with 1mL TBS buffer, 10 min nutation between washes after last wash, removed TBS, quick spin, remove remaining TBS with P20 added 100ul elutiion buffer to each sample Added 50ul elution buffer to inputs put at 65deg for 8 hours Washed IgG bead samples after 3 hour nutation at 4deg, 6x with RIPA buffer, 10min nutation between each wash washed twice with TE pH 7.6, 3 minute nutation between washes removed TE after last wash, quick spin, removed remaining TE buffer with P20 added 100ul elution buffer to each sample put at 65deg for 5 hours 30 mins at 37deg with 1ul RNase A 2 hours at 62deg with 0.5ul proteinase K purify with Sigma columns 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