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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
into E. coli DH5-alpha.
Inoculum of
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 I34 insert bands were 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)
September, 15th
September, 16th
September, 17th
September, 18th
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