Team:Arizona State/Notebook/hyder


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Hyder's lab notes


miniprepped gfp1 and rbs1 and rbs2 liquid cultures

picked 1 colony from double terminator (dt1) plate

picked 1 colony from t7 polymerase (pol1) plate

picked 1 colony from puc19 plate (positive control)

picked 1 colony from dh5a plate (negative control)

started liquid cultures of each colony (5 mL LB amp each)


Resuspended GFPT1 and GFPT2 oligos with molecular grade (nuclease-free) H2O.

Final Concentration 100uM

(gfpt1 top1, gfpt2 top1, gfpt1 top2, gftp2 top2, gfpt1 bot1, gfpt2 bot1, gfpt1 bot2, gfpt2 bot2)

(3uL of each oligo + 2uL 10x annealing buffer, 6uL molecular grade H2O. 20uL Reactions)

Heated for 5 minutes at 100C. Let cool to room temperature on the heating block, stored at -20C.

Digested BBa_I13522 with XbaI and PstI.

Attempted ligating annealed oligos into a digested ?kill plasmid? from Ryan (realized it was cut with E and P).


Annealed oligos for GFPT1 and GFPT2 (target probes)

Ligated oligos with digested GFP plasmid (BBa_I13522)

Transformed into competent DH5alpha

Added SOC and incubated at 37C for 15 minutes.

Plated on amp treated plates.


Only one colony on each plate (both were white)

Picked colonies and started 5mL LB amp cultures of each, stored at 37C

Stored plates in 37C