Terebratalia Vivo-Morpholino engrailed/wnt1 with permeabilization

Use 25 µM concentration for vivo-morpholinos.

CTRL FSW
CTRL CFSW 1h Ttra_en_TB_vMO 25 µM Ttra_en_SBe2i2_vMO 25 µM Ttra_wnt1_TB_vMO 25 µM Ttra_wnt1_SBe2i2_vMO 25 µM
CTRL Pronase 10 µg/mL 1h Ttra_en_TB_vMO 25 µM Ttra_en_SBe2i2_vMO 25 µM Ttra_wnt1_TB_vMO 25 µM Ttra_wnt1_SBe2i2_vMO 25 µM
CTRL Electroporation in CFSW Ttra_en_TB_vMO 25 µM Ttra_en_SBe2i2_vMO 25 µM Ttra_wnt1_TB_vMO 25 µM Ttra_wnt1_SBe2i2_vMO 25 µM

Electroporation setup

BioRad Gene Pulser 50V, 25 µF, resistance set to infinity.

[photo]

ms
ctrl 1.1
en tb 1.0
en sb 1.1
wnt1 tb 1.2
wnt1 sb 0.9

16/01/14

  • Calcium-free seawater look normal trilobed.
  • Pronase treatment embryos failed to gastrulate, including the control.
  • Electroporation look normal except for wnt1 sb which seem either delayed or did not manage to close the blastopore.