Probe synthesis of Novocrania Hedgehog pathway

Selected minipreps for probe synthesis of Novocrania Hedgehog pathway and remaining Terebratalia genes. From this post.

BV326 Terebratalia transversa Lmx1 T7 ok + SP6
BV328 Terebratalia transversa NK1a T7 ok + SP6
BV330 Terebratalia transversa NK1b T7 ok + SP6
BV332 Novocrania anomala Patched1 T7 ok + SP6
BV336 Novocrania anomala Smoothened1 T7 ok + SP6
BV338 Novocrania anomala Smoothened2 T7 ok + SP6
BV340 Novocrania anomala Gli T7 ok + SP6
BV343 Novocrania anomala Pax6 T7 ok + SP6
BV345 Novocrania anomala Pax3/7 T7 ok – T7
BV346 Novocrania anomala NK1 T7 ok + SP6
TTR Terebratalia Pax6 – – – T7 DIG
TTR Terebratalia Pax6 – – – T7 DNP

Set probe PCR to run overnight.

19/11/14

Ran gel and painfully extracted the PCR…

2014-11-19 19.09.46

20/11/14

Started probe synthesis for the genes above at 9:30. Finished at 16:30 and precipitated overnight.

21/11/14

gene ng/µL goal concentration volume total volume volume of hybe buffer
Ttra lmx1 906 50 24 426 402
Ttra nk1a 974 50 24 458 434
Ttra nk1b 1098 50 24 516 492
Nano ptc1 1075 50 24 505 482
Nano smo1 2041 50 24 959 936
Nano smo2 1830 50 24 860 836
Nano gli 488 50 24 229 206
Nano pax6 1350 50 24 634 611
Nano pax37 2224 50 24 1045 1022
Nano nk1 1050 50 24 494 470
Ttra pax6 dig 1076 50 24 506 482
Ttra pax6 dnp 50 50 24 23 0