Novocrania one more time

Using Andi’s primers with annealing temperature at 57 °C.

11/09/13

Engrailed rendered a band at the expected size (638 bp), although it is a bit spread band. It is quite possible that wnt1 had a band at the right place (718 bp), but it was very weak; I cut it anyway. Wnt5 band was also at the correct place (932 bp), but also weak.

2013-09-11 16.37.23

Set ligation reaction at 16h and stored at 4°C.

12/09/13

Transformation and plating.

13/09/13

en had a lot of colonies, wnt1 had a normal amount, and wnt5 had a few colonies. I managed to do 10 colonies for the first two and 8 colonies for wnt5:

2013-09-16 10.57.37

16/09/13

Picked colonies for minipreps

gene colony T7
en 8 BV266
en 9 BV267
en 10 BV268
wnt1 1 BV269
wnt1 2 BV270
wnt1 3 BV271
wnt5 1 BV272
wnt5 2 BV273
wnt5 3 BV274

17/09/13

  • Extracted plamids.

18/09/13

  • Set sequencing pcr at 10:30.
  • Dropped sequences at the facility at 14:30.

22/09/13

Checked the alignment and it looks good! See files: Nano_en-wnt1-wnt5