Lab Solutions and Reagents: Difference between revisions
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:PTw Containing 4% Paraformaldehyde | :PTw Containing 4% Paraformaldehyde | ||
*'''[[DNA/RNA preparation]]''' | *'''[[DNA/RNA preparation]]''' | ||
:10x mRNA Transcription Buffer | |||
:2x mRNA Nucleotide Triphosphate Mix | |||
:TE | |||
:Homoginization Buffer | |||
:Reticulocyte Standard Buffer (RSB) | |||
:Nuclei Freezing Buffer | |||
*'''[[embryo manipulation/dissection]]''' | *'''[[embryo manipulation/dissection]]''' | ||
:Agarose Plates, 1% (for use with trops or laevis) | :Agarose Plates, 1% (for use with trops or laevis) | ||
| Line 66: | Line 72: | ||
:Rehydrating Embryos(for embryos stored in 100% methanol) | :Rehydrating Embryos(for embryos stored in 100% methanol) | ||
*'''[[microinjection]]''' | *'''[[microinjection]]''' | ||
:Ficoll, 2.5% | |||
:2.5% Ficoll in 1/3X MR (for use with X. laevis embryos) | |||
:3% Ficoll in 1/9XMR (for use with X.tropicalis embryos) | |||
*'''[[general buffers and salt solutions]]''' | *'''[[general buffers and salt solutions]]''' | ||
:HEPES 1 M, pH 7.4 | :HEPES 1 M, pH 7.4 | ||
| Line 79: | Line 88: | ||
:1M Tris pH 9.5 | :1M Tris pH 9.5 | ||
'''Additional Solutions/Reagents:''' | |||
*[[benzocaine stock, 100x (Khokha lab)]] | *[[benzocaine stock, 100x (Khokha lab)]] | ||
*[[high-salt solution for egg laying (Zorn lab/Ron Laskey)]] | *[[high-salt solution for egg laying (Zorn lab/Ron Laskey)]] | ||
*[[dejellying solution (Zorn lab)]] | |||
*[[recipes for miscellaneous salts, solutions, etc. (Khokha lab)]] | *[[recipes for miscellaneous salts, solutions, etc. (Khokha lab)]] | ||
Latest revision as of 12:54, 16 December 2011
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).
Additional methodology adapted from Khokha lab protocols [2]
click each bulleted list heading to view 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
- MEMFA
- Dent's Fix (Methanol/DMSO)
- Bleaching solution (Mayor et al. 1995)
- Goat Serum (GS)
- PBT
- DAB
- H2O2
- Secondary Antibodies Coupled to Horseradish Peroxidase
- Murray's Clearing Medium
- Nucleotide Stocks
- 5x SP6 Buffer
- MEMFA
- PTw
- Triethanolamine 0.1 M
- PTw Containing 4% Paraformaldehyde
- Hybridization Buffer
- SSC, 20X
- Denhardt’s solution, 100X
- RNase A
- RNase T1
- 4% BMB Blocking Reagent in MAB
- Alkaline Phosphatase Buffer
- Levamisol 1 M
- NBT/BCIP
- PVA Alkaline Phosphatase Buffer
- Bouin’s Fixative
- Bleaching solution (Mayor et al. 1995)
- MEMFA
- Bouin’s Fixative
- Dent's Fix (Methanol/DMSO)
- Paraformaldehyde(PFA), 20%
- PTw Containing 4% Paraformaldehyde
- 10x mRNA Transcription Buffer
- 2x mRNA Nucleotide Triphosphate Mix
- TE
- Homoginization Buffer
- Reticulocyte Standard Buffer (RSB)
- Nuclei Freezing Buffer
- Agarose Plates, 1% (for use with trops or laevis)
- Dehydrating embryos
- Rehydrating Embryos(for embryos stored in 100% methanol)
- Ficoll, 2.5%
- 2.5% Ficoll in 1/3X MR (for use with X. laevis embryos)
- 3% Ficoll in 1/9XMR (for use with X.tropicalis embryos)
- HEPES 1 M, pH 7.4
- 1M MgCl2
- 10x Phosphate Buffered Saline (PBS)
- 20x Phosphate Buffered Saline (PBS)
- PBT
- Proteinase K(PK), 10 mg/ml
- PTw
- 4M NaCl, 1 liter
- SDS 10%
- TAE 50X
- 1M Tris pH 9.5
Additional Solutions/Reagents: