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.


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

2013-09-23 23.37.45

Extracted the probe PCR.


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


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.


Good engrailed concentration! 1334 ng/µL.