Skip to main content

Table 2 Reagents

From: Simultaneous high-resolution detection of multiple transcripts combined with localization of proteins in whole-mount embryos

Reagents Composition Provider
Danieau’s solution 17.4 mM NaCl, 0.21 mM KCl, 0.12 mM MgSO4•7H2O, 0.18 mM Ca(NO3)2, 1.5 mM Hepes  
Fixation solution 4% PFA in PBS, pH 7  
PBS 137 mM NaCl, 2.7 mM KCL, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4  
PBT 0.1%/0.01% 0.1%/0.01% Tween-20 in PBS  
SSCT 0.01% 0.01% Tween-20, 15 mM NaCl, 1.5 mM TriNaCitratdihydrate, pH 7  
Negative control dapB (cat.# 310043) Channel 1, ready-to-use solution Advanced Cell Diagnostics (ACD), Hayward, CA, USA
Target probe cxcl12a (cat.# 406481) Channel 1: ready-to-use solution ACD, Hayward, CA, USA
Target probe myoD (cat.# 402461-C2) Channel 2: 50× solution ACD, Hayward, CA, USA
Target probe vasa (cat.# 407271-C3) Channel 3: 50× solution ACD, Hayward, CA, USA
Target probe cxcr4b (cat.# 418121-C3) Channel 3: 50× solution ACD, Hayward, CA, USA
Target probe egfp (cat.# 400281) Channel 1: ready-to-use solution ACD, Hayward, CA, USA
Target probe nanos (cat.# 404521-C2) Channel 2: 50× solution ACD, Hayward, CA, USA
Pretreat 3 in RNAscope pretreatment kit (cat.# 320842) Pretreat 3 ready-to-use solution ACD, Hayward, CA, USA
RNAscope fluorescent multiplex detection kit (cat.# 320851) Amp1, Amp2, Amp3, Amp4 AltA, Amp4 AltB, Amp4 AltC, Pretreat 3, DAPI ready-to-use solutions, Wash buffer provided in the kit is not used and is replaced with 0.2× SSCT (0.01% Tween-20) ACD, Hayward, CA, USA
1% or 4% LMP 1% or 4% low melting point agarose in Danieau’s solution Invitrogen, Paisley, UK
Hoechst 33342 (cat.# H1399) Use at a 1:10000 dilution in 0.2× SSCT (0.01% Tween-20) Invitrogen, Paisley, UK
  1. PBS, phosphate-buffered saline; PBT, PBS buffer + Tween-20; PFA, paraformaldehyde; SSCT, saline-sodium citrate buffer + 0.01% Tween-20.