Brachiopod cloning

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Began PCR for cloning Novocrania en, wnt1, wnt5 and Terebratalia fgfr, lfng, pax2/5/8, Fz2, Fz4, Fz5/8, Fz7, Fz10, aldh2, arm.


Gel, Novocrania genes did not work. Others did, extraction and ligation.

2013-08-01 15.09.57

I started a PCR for Novocrania genes using a lower annealing temperature of 55 °C.


Managed one band for engrailed, but not at the right length, slightly shorter.

2013-08-02 18.29.52


I reset the ligation for: lfng, fz58 7, aldh2, arm.

Colony PCR running for the rest.

Also started a PCR with Nano wnt1 and wnt5 with annealing temperature of 50 °C.


Transformed and plated the genes that did not work previously (lfng, fz5/8, fz7, aldh2, arm).

Novocrania gel is not promising, bands are not the right size, need to design new primers.

2013-08-05 13.24.23

Colony PCR went well and today I’ll put the colonies to grow.

2013-08-05 13.24.13

Animal Gene Colony T7
Ttra FGFR 1 BV248
Ttra FGFR 2 BV249
Ttra FGFR 3 BV250
Ttra Pax 2/5/8 1 BV251
Ttra Pax 2/5/8 2 BV252
Ttra Pax 2/5/8 3 BV253
Ttra Fz2 1 BV254
Ttra Fz2 2 BV255
Ttra Fz2 3 BV256
Ttra Fz4 2 BV257
Ttra Fz4 3 BV258
Ttra Fz10 2 BV259
Ttra Fz10 3 BV260
Ttra Fz10 4 BV261
Nano En 3 BV262


Sequences above dropped at the sequencing facility after miniprep extraction and sequencing pcr.

Also, set colony pcr for the genes that hadn’t worked.


Colony gel, putting good colonies to grow today.

2013-08-07 12.43.32


Extracted minipreps for Lfng 3 and 5 and Aldh2 1. Sequencing PCR and dropped samples in the facility.

Gene ID
LFNG c3 BV263
LFNG c5 BV264
ALDH2 c1 BV265