Lab Solutions and Reagents: Difference between revisions

From XenWiki
Jump to navigation Jump to search
imported>Virgilio
No edit summary
imported>Virgilio
No edit summary
Line 85: Line 85:


*[[recipes for miscellaneous salts, solutions, etc. (Khokha lab)]]
*[[recipes for miscellaneous salts, solutions, etc. (Khokha lab)]]
: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

Revision as of 09:09, 14 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
Agarose Plates, 1% (for use with trops or laevis)
Dehydrating embryos
Rehydrating Embryos(for embryos stored in 100% methanol)
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