Team:UNIPV-Pavia/Calendar/September/settimana3

From 2010.igem.org

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==September, 14th==
==September, 14th==
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Finally Mr.Gene sent us our UPDATED_YEAST: a little difficult to pick up the package but at the end we succeeded!
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Finally Mr.Gene sent us our YEAST: a little difficult to pick up the package but at the end we succeeded!
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Revision as of 20:27, 14 September 2010


SEPTEMBER: WEEK 3



September, 13th

Unfortunately we discovered we tried to ligate I32 (E-S) to I37 (X-P) instead of I37 (E-X) ;-(

So this step will be repeated.

Trasformation of ligations

  • I52
  • I54
  • I55
  • I56
  • I57
  • I58

into E. coli DH5-alpha.

Inoculum of

  • I26
  • I31
  • I34

into 5 ml LB+Amp for ligations of the following day.



September, 14th

Finally Mr.Gene sent us our YEAST: a little difficult to pick up the package but at the end we succeeded!


I52, I54, I55, I56, I57, I58 plates showed in general few colonies; I52 and I5X showed very few colonies (<=5). Very strange (mumble mumble...), they will be screened ASAP (stored at +4°C).

Miniprep and quanfification of:

  • I26: 69,7 ng/ul
  • I31: 89,5 ng/ul
  • I34: 147,6 ng/ul

3-hours digestion:

  • I26: XbaI-PstI (Insert)
  • I31: EcoRI-SpeI (Insert)
  • I34: E-X (Vector)

Gel run/cut of samples (I26 and I31 insert bands were very very soft)

Gel run/cut for digested I26, I31 and I34.

and gel extraction:

  • I26 (X-P): 2,6 ng/ul
  • I31 (E-S): 1,4 ng/ul
  • I34 (E-X): ng/ul

We already had digested DNA so we could perform new ON ligations

  • I59: I21 (E-S) + I34 (E-X)
  • I60: I20 (S-P) + I26 (X-P)
  • I61: I31 (E-S) + I37 (E-X)

and repeat

  • I53: I32 (E-S) + I37 (E-X)


Inoculum of I47, I48, I49 into 5 ml LB+Amp for TECAN test of the following day.



September, 15th

I47, I48, I49 cultures were diluted 1:100 into 5ml LB+Amp. In the afternoon all cultures were synchronized to O.D. 0,02 and TECAN test was started to check GFP production by these BioBricks.

September, 16th

September, 17th

September, 18th