Team:UNIPV-Pavia/Calendar/September/settimana3

From 2010.igem.org

(Difference between revisions)
(September, 14th)
m (September, 14th)
Line 66: Line 66:
*I34: ng/ul
*I34: ng/ul
3-hours digestion:
3-hours digestion:
-
*I26: XbaI-PstI
+
*I26: XbaI-PstI (Insert)
-
*I31: EcoRI-SpeI
+
*I31: EcoRI-SpeI (Insert)
-
*I34: E-X
+
*I34: E-X (Vector)
Gel run/cut of samples
Gel run/cut of samples
[[Image:UNIPV10_14_09_10_gel_run_cut_phasins_2setp_final.jpg|thumb|200px|center| Gel run/cut for I26, I31 and I34.]]
[[Image:UNIPV10_14_09_10_gel_run_cut_phasins_2setp_final.jpg|thumb|200px|center| Gel run/cut for I26, I31 and I34.]]

Revision as of 19:38, 13 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

Miniprep and quanfification of:

  • I26: ng/ul
  • I31: ng/ul
  • I34: ng/ul

3-hours digestion:

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

Gel run/cut of samples

and gel extraction:

  • I26 (X-P): ng/ul
  • I31 (E-S): ng/ul
  • I34 (E-X): ng/ul

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

  • IXX: I32 (E-S) + I37 (E-X)
  • IXX: I21 (E-S) + I34 (E-X)
  • IXX: I31 (E-S) + I37 (E-X)

and repeat

  • I53: I32 (E-S) + I37 (E-X)

September, 15th

September, 16th

September, 17th

September, 18th