Team:UNIPV-Pavia/Calendar/September/settimana3

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Revision as of 19:33, 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

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

September, 16th

September, 17th

September, 18th

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