Team:UNIPV-Pavia/Calendar/September/settimana3

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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

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

and gel extraction:

  • I26 (X-P): 69,7 ng/ul
  • I31 (E-S): 89,5 ng/ul
  • I34 (E-X): 147,6 ng/ul

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

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

and repeat

  • I53: I32 (E-S) + I37 (E-X)


September, 15th

September, 16th

September, 17th

September, 18th