IGEM:MIT/2006/Notebook/2006-8-12

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osmY fluorescence test

Put 3 1:500 dilutions of each of the following full-grown cultures into the plate reader:

R0040.E0840 (wells G1, G2, and G3)

osmY.E0840 A (wells G4, G5, and G6)

osmY.E0840 A GE (wells G7, G8, and G9)

osmY.E0840 C (wells G10, G11, and G12)

osmY.E0840 CS (wells H1, H2, and H3)

Made LCs of the following transformants

30.BAT2 C mut (A, B, and C) (AMP)

30.BAT2 B mut (A, B, and C) (AMP)

200 mut (A, B, and C) (AMP)

Q04400.J45119 (A, B, and C) (AMP/TET)

Q04400.J45199 (A, B, and C)* (AMP/TET)

Q04400.J4549 (A, B, and C)** (AMP/TET)

THI3 (A, B, and C)*** (AMP)

BB pCHBA A (A, B, and C) (AMP)

Q04400.J45099 (A, B, and C) (AMP/TET)

pUCP22+J45100 (A, B, and C) (AMP/GEN)

pUCP22+J45120 (A, B, and C) (AMP/GEN)

BB pCHBA A/K (A, B, and C) (AMP/KAN)

32.BAT2 mut (A and B)**** (AMP)

220 mut (A, B, and C) (AMP)

BAT2 mut did not transform

osmY.E0840 did not transform

Note: 250 A mut, 250 B mut, 250 C mut, 270 B mut, and 270 C mut were not LCed since their original sequencing results were incorrect.

Additional Note: Both pUC18 plates had lots of transformants on them.

One stars: Actually labeled "Q04400+J451199" (I just assumed that it was J45199, not J451199).

Two stars: Do not know what J4549 means (I continued to label it in the same manner).

Three stars: All of these colonies were very small although I am pretty sure they're not just bubbles.

Four stars: Only two transformants, which ran into each other. I will try to extract from the opposite edge of each colony.