Part:BBa_K1129006:Experience

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1129006

The Cas9 construct was transformed into E. cloni 10G, plated on LB agar plates made with chloramphenicol (33ug/mL), and grown overnight at 37C.

A single colony was picked and grown overnight in LB (with chloramphenicol) at 30C and shaking (1000 rpm). Growth was very slow for the Cas9 culture and OD was only ~0.6 in the morning. Several aliquots of 175 uL were taken and placed in separate wells of a 96-well plate. They were induced with varying concentrations of arabinose (0, 0.5%, 1%, and 2%) for three hours at 37C and 1000 rpm.

OD was measured and it was determined that there was a tradeoff between induction concentration and growth. The highest induction concentration that also had relatively good growth was 1% arabinose (OD=1.17 vs 0.98 for 2% induction and 1.67 for non-induced). This culture and the non-induced control was collected, centrifuged, resuspended in SDS-PAGE loading buffer, and boiled for 10 minutes. They were loaded in a

[File:UBC-Growth_curves_MOI.png|800px]

User Reviews

UNIQ6e509c0a465ceb97-partinfo-00000001-QINU UNIQ6e509c0a465ceb97-partinfo-00000002-QINU