Probe synthesis of Novocrania segmentation

After successfully cloning Novocrania genes using Andi’s primers it is time for probe synthesis.

Set Probe PCR for en, wnt1 and wnt5.

23/09/13

Gel and extraction went fine, except that I forgot the dna ladder…

2013-09-23 23.37.45

Extracted the probe PCR.

24/03/13

Began probe synthesis transcription reaction at 10h and stopped at 16h. Everything normal.

25/03/13

After centrifuging there was a very small pellet for engrailed. Wnt1 and wnt5 had normal pellets. Here are the nanodrop:

Plasmid Gene Enzyme ng/µL
BV267 engrailed SP6 13.08
BV271 wnt1 SP6 156.92
BV274 wnt5 SP6 54.99

Since engrailed was so low I started a new transcription reaction at 12h.

26/09/13

Good engrailed concentration! 1334 ng/µL.