Lab Solutions and Reagents: Difference between revisions

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*[[culture media]]
'''most information adapted from:'''
 
''Early Development of Xenopus Laevis: A Laboratory Manual.''[http://www.xenbase.org/literature/book.do?method=display&bookId=22]
 
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 [http://khokha.medicine.yale.edu/protocols_general/protocols_general.html]
 
 
'''click each bulleted list heading to view solution/reagent recipes'''
 
*'''[[culture media]]'''
:Modified Barth's Saline (MBS)
:Modified Barth's Saline (MBS)
:Anitbiotics
:Anitbiotics
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:Normal Amphibian Medium (NAM)
:Normal Amphibian Medium (NAM)
:Holtfreter's Solution
:Holtfreter's Solution
*[[immunohistochemistry]]
*'''[[immunohistochemistry]]'''
*[[in situ hybridization]]
:MEMFA
*[[fixatives]]
:Dent's Fix (Methanol/DMSO)
*[[DNA/RNA preparation]]
:Bleaching solution (Mayor et al. 1995)
*[[embryo manipulation/dissection]]
:Goat Serum (GS)
*[[microinjection]]
:PBT
*[[general buffers and salt solutions]]
:DAB
:H2O2
:Secondary Antibodies Coupled to Horseradish Peroxidase
:Murray's Clearing Medium
*'''[[in situ hybridization]]'''
: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)
*'''[[fixatives]]'''
:MEMFA
:Bouin’s Fixative
:Dent's Fix (Methanol/DMSO)
:Paraformaldehyde(PFA), 20%
:PTw Containing 4% Paraformaldehyde
*'''[[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]]'''
:Agarose Plates, 1% (for use with trops or laevis)
:Dehydrating embryos
:Rehydrating Embryos(for embryos stored in 100% methanol)
*'''[[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]]'''
: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:'''


*[[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)]]
**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

Latest revision as of 13:54, 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:


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