Anesthesia solution |
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Serumwerk Bernburg GmbH (Ursotamin), Bayer Health Care (Rompun) |
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Mixture of 1 ml Ursotamin (100 mg/ml Ketaminhydrochlorid) and 240 μl Rompun (2% Xylazinhydrochlorid) |
Citrate solution |
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117,64 mg sodium citrate in 10 ml 0.9% NaCl solution |
Solution A |
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134 mM NaCl, 4 mM KCl, 11 mM glucose, 1.2 mM MgSO4, 1.2 mM Na2HPO4, 10 mM HEPES, pH adjusted to 7.35 using 10 M NaOH, sterile filtered |
Solution A+ |
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Content of solution A plus 0.2 mM EGTA, pH adjusted to 7.35 using 10 M NaOH, sterile filtered |
Liberase solution |
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F. Hoffmann‐ La Roche Ltd. |
11988476 001 |
500 μl stock solution in 15 ml solution A (stock solution:
10 mg/ml Liberase Blendzyme 4 in aqua dest.) |
Solution B |
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Content of solution A plus 200 μM CaCl2 and 0.1% DNase solution |
Solution B1/2 |
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1:1 mixture of solution A and solution B |
DNase solution |
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Sigma‐ Aldrich Co. |
D4527 |
Deoxyribonuclease I, Type 2 from bovine pancreas, 40 KU (15 mg) are dissolved in 5 ml of 10 mM Tris‐HCl buffer, adjusted to pH=7.35, additional content: 50 mM NaCl, 10 mM MgCl2, 1 mM dithioerythritol; this solution is mixed with 5 ml glycerol |
Extracellular matrix protein (ECM) solution |
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Extracellular matrix protein (ECM) solution |
E1270 |
1 ml stock solution as purchased is diluted with 6 mL
medium M199 |
Culture medium M199 |
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PAA |
E15‐834 |
Medium M199 supplemented with 1 μl/ml ITS solution and 20 μl/ml penicillin‐streptomycin solution (5000 units/ml) |
ITS solution |
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25 mg Insulin, 25 mg Transferrin and 50 μl Selenite stock solution are dissolved in 5 ml aqua dest. (stock solution: 0.5 mg/ml Sodiumselenite in aqua dest.). Add a few drops of 1 M HCl until one should get a clear
solution. Aliquots can be frozen. |
Tyrode |
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135 mM NaCl, 5.4 mM KCl, 1.8 mM CaCl2, 2mM MgCl2, 10 mM glucose, 10 mM HEPES, pH adjusted to 7.35 using 10 M NaOH, sterile filtered |
Master mix |
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5 μl 10×PCR‐Buffer, 5 μl MgCl2 (25 mM), 2 μl forward primer, 2 μl reverse primer, 1 μl dNTP´s (10 mM each), 33 μl H2O, 1 μl DMSO, 1 μl Taq‐DNA polymerase for each sample |
LB‐Medium |
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10 g/l Tryptone, 5 g/l Yeast extract, 5 g/l NaCl (and 15 g/l Agar if used as solid medium), pH 7.0 |
Pac I |
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New England Biolabs |
R0547L |
preparation: 50 μg DNA, 5 μl 10×buffer, 0.5 μl 100×BSA,
0.75 μl Pac I, fill with H2O up to final volume of 50μl |