In situ Hybridization of Embryos with Antisense RNA Probes - Caenorhabditis Elegans: Molecular Genetics and Development - page 260

In our hands, PVA in the developer greatly improves signal quality, and is worth
the trouble to make.
14. Developer Solution

2 TNM 25 mL

2 PVA 25 mL

NBT 200 m L

BCIP 200 m L

Levamisole 7.2 mg

Mix reagents in a Falcon tube and mix by inversion. Prepare immediately before use.

VII. Notes

A. Suggested Positive Controls

Gene Expression Pattern Primers (forward primer, T7-tagged reverse primer)

5 0 -gtt caaggaatcaactcgaag act c-3 0 ,5 0 -TAATAC
GAC TCA CTATAG GGA CTtggcagagct
act gat ttcgtt gga-3 0

pgl-1 Germ line and

embryos

5 0 -ttc aatcgtacgatc cag cacaacaat cg-3 0 ,5 0 -TAA
TAC GAC TCA CTATAG GGA CTT GCA CT
caatagctcctgaatcagt t-3 0

end-1 Early E lineage

(E, Ea/Ep)

5 0 -aaaccagccagcttactacctcccgtccta-3 0 ,5 0 -
TAATACGACTCACTATAGGGACTcgttcccga
gcttatgatgatctctatc-3 0

hlh-1 Mid to late

embryogenesis,
muscle lineages

5 0 -gtaatggcgattggactctcacatatgacc-3 0 ,5 0 -
TAATACGACTCACTATAGGGACTtctgctgccg
tgatgacaacgatttgtg-3 0

opt-2 Mid to late

embryogenesis,
intestine

5 0 -gga tgttgtccaaga gat gaatccacc a-3 0 ,5 0 -TAA
TAC GAC TCA CTATAG GGA CTtgttcaa tat
cgcaagaagcgacac gtc-3 0

myo-2 Late stage embryos,

pharynx muscle

5 0 -tct cgtgtcgtccgt cag gcaccagga gag-3, 5 0 -TAA
TAC GAC TCA CTATAG GGA CTcctggtgatg
atccacttgaacata cgg-3 0

myo-3 Late stage embryos,

body muscle

5 0 -gtagtagcatctctaatatccatgacgttc-3 0 ,5 0 -
TAATACGACTCACTATAGGGACTtccttgaac
gtattaggaatgtattctt-3 0

cup-4 4 coelomocytes in

embryos, 6 in
adults