Revision as of 22:35, 2 July 2009

Week from July 1st, to July 5th, 2009

Miniprep for:
- A8 (for dilution/transformation)
- A9 (for dilution/transformation)

We transformed 10 pg of A8 and A9 miniprepped DNA in TOP10, after a proper quantification with Nanodrop and dilution. We plated transformed bacteria and incubated them overnight at 37°C.

We infected 5 ml of LB + Amp with 10 ul of T9002 glycerol stock. We incubated the culture overnight at 37°C, 220 rpm.

We also picked a random colony from A10 plate and infected 1 ml of LB + Amp. We incubated this inoculum at 37°C, 220 rpm for 5 and 1/2 hours. Then we prepared a glycerol stock and re-filled the remaining 250 ul of culture with 5 ml of LB + Amp to grow an overnight culture (for sequencing).

Preparation of experiment with Tecan F200

Experiment with Tecan F200

Miniprep for A10 and T9002. We sent purified DNA to BMR Genomics for sequencing.

A8(10pg) and A9(10pg) plates showed colonies!(a few, as expected) We picked 3 colonies for A8 plate and 3 colonies for A9 plate to infect 5 ml of LB + Amp. We incubated the six inocula at 37°C, 220 rpm overnight. Tomorrow they will be miniprepped and screened with digestion to check if the extra band had been lost!

Preparation of experiment with Tecan F200

We incubated the diluted cultures at 37°C, 220 rpm for 3 hours. After 2 hours, one of the two A9 culture was induced with 75 ng/ml aTc and incubated again under the same conditions as before.

Experiment with Tecan F200

@@ Line 69: / Line 69: @@
 *Miniprep for A10 and T9002. We sent purified DNA to BMR Genomics for sequencing.
+*A8(10pg) and A9(10pg) plates showed colonies!(a few, as expected) We picked 3 colonies for A8 plate and 3 colonies for A9 plate to infect 5 ml of LB + Amp. We incubated the six inocula at 37°C, 220 rpm overnight. Tomorrow they will be miniprepped and screened with digestion to check if the extra band had been lost!
 ''Preparation of experiment with Tecan F200''
+*aTc stock preparation.
 *We diluted 1:1000 the cultures of:
 **A8
-**A9
+**A9 (X2)
 **E0240
-*We incubated the diluted cultures at 37°C, 220 rpm for 3 hours.
+*We incubated the diluted cultures at 37°C, 220 rpm for 3 hours. After 2 hours, one of the two A9 culture was induced with 75 ng/ml aTc and incubated again under the same conditions as before.