Lab Solutions and Reagents
most information adapted from:
Early Development of Xenopus Laevis: A Laboratory Manual.[1]
2000; (First ed.). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
Sive, Hazel (Author), Grainger, Robert M (Author), and Harland, Richard M (Author).
please click each bulleted list heading to see solution/reagent recipes
- Modified Barth's Saline (MBS)
- Anitbiotics
- Gentamycin
- Oxytetracycline
- Penicillin
- Streptomycin Sulfate
- High-salt MBS
- Amphibian Ringers
- Modified Frog Ringers (MR)
- Marc's Modified Ringer's (MMR)
- Normal Amphibian Medium (NAM)
- Holtfreter's Solution
- immunohistochemistry
- in situ hybridization
- fixatives
- DNA/RNA preparation
- embryo manipulation/dissection
- microinjection
- general buffers and salt solutions
- agarose plates 1% - for use with laevis or tropicalis
- Alkaline Phosphatase (AP) Buffer, with Levamisol and Tween
- amphibian Ringers, 5 gal (19 L)
- bleaching solution (Sive et al. 2000, Mayor et al. 1995)
- 4% BMB blocking reagent in MAB
- Bouin’s fixative
- cysteine, 2.0%
- dehydrating embryos
- Denhardt’s solution, 100X
- dNTPs 10 mM
- 2.5% Ficoll
- 2.5% Ficoll in 1/3X MR (for use with X. laevis embryos)
- 3% Ficoll in 1/9XMR (for use with X.tropicalis embryos)
- gentamicin stock, Sigma (1000X, 100 mg/ml)
- HEPES 1 M, pH 7.4
- hybridization buffer
- levamisol 1 M
- MAB, 10X
- 1M MgCl2
- MEMFA* (Sive et al. 2000)
- 100X MR salts
- 1/3X Modified Ringers
- 1/9X MR + 1% Gentamicin
- 1/20X MR
- modified Ringers
- paraformaldehyde(PFA), 20%
- phosphate buffered saline (PBS), 10X
- proteinase K(PK), 10 mg/ml
- PTw
- rehydrating embryos (for embryos stored in 100% methanol)
- RNase A (Sigma R 5000)
- RNase T1 (Sigma R8251)
- 4M NaCl, 1 liter
- SDS 10%
- Sodium Phosphate Buffer(1M)
- SSC, 20X
- TAE 50X
- Triethanolamine 0.1 M (pH7.0-8.0)
- 1M Tris pH 9.5