User:Perry/Spring 2007: Difference between revisions
(New page: ==2/12/07== dissolved 500mg ampicillin salt into 10ml water to prepare 50mg/ml ampicillin stock; aliquotted 1ml into tubes, left in -20dC. overnight 1ml LB-amp cultures of JLO159, W, H, S...) |
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159- .335, W- .245, H- .231, S- .279 | 159- .335, W- .245, H- .231, S- .279 | ||
made liquid cultures of 5ml LB-amp | made liquid cultures of 5ml LB-amp + 250ul overnight cultures. incubated 3h, added 5mM IPTG (53ul 0.5M), incubated 2h, diluted 1:10, measured OD600. | ||
aliquotted volumes into tubes. centrifuged 8000rpm, 3min. aspirated. reconstituted in 10ul streptavidin aptamer, 10ul B/F hybrid, or 10ul B0/F hybrid. | |||
4uM streptavidin aptamer: 48ul PBS + 2ul streptavidin aptamer (100uM) | |||
4uM B/F hybrid: 46ul PBS + 2ul Bside oligo (100uM) + 2ul Iside oligo (100uM) | |||
4uM B0/F hybrid: 46ul PBS + 2ul DispI oligo (100uM0 + 2ul Iside oligo (100uM) | |||
Wrapped tubes in a napkin, rotated on rotisserie at room temperature for 1h. | |||
Revision as of 12:40, 13 February 2007
2/12/07
dissolved 500mg ampicillin salt into 10ml water to prepare 50mg/ml ampicillin stock; aliquotted 1ml into tubes, left in -20dC.
overnight 1ml LB-amp cultures of JLO159, W, H, S, ~4:30pm.
2/13/07
took overnight cultures out ~7:30pm. diluted 1:10, measured OD600.
159- .335, W- .245, H- .231, S- .279
made liquid cultures of 5ml LB-amp + 250ul overnight cultures. incubated 3h, added 5mM IPTG (53ul 0.5M), incubated 2h, diluted 1:10, measured OD600.
aliquotted volumes into tubes. centrifuged 8000rpm, 3min. aspirated. reconstituted in 10ul streptavidin aptamer, 10ul B/F hybrid, or 10ul B0/F hybrid.
4uM streptavidin aptamer: 48ul PBS + 2ul streptavidin aptamer (100uM)
4uM B/F hybrid: 46ul PBS + 2ul Bside oligo (100uM) + 2ul Iside oligo (100uM)
4uM B0/F hybrid: 46ul PBS + 2ul DispI oligo (100uM0 + 2ul Iside oligo (100uM)
Wrapped tubes in a napkin, rotated on rotisserie at room temperature for 1h.
