Team:Arizona State/Notebook/hyder

(Difference between revisions)

Revision as of 07:44, 1 October 2012

Hyder's lab notes

6/22/12

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)

8/3/12

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).

8/6/12

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.

8/7/12

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

8/8/12

@@ Line 31: / Line 31: @@
+Digested BBa_I13522 with XbaI and PstI.
-BBa_I13522
+Attempted ligating annealed oligos into a digested ?kill plasmid? from Ryan (realized it was cut with E and P).
+/6/12
+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.
+/7/12
+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
+/8/12