Crosley CRSH268MW2 Bedienungsanleitung PDF herunterladen (Seite 16)

3 Material and Methods

Media / Antibiotics Composition

LB -Medium 1 % Tryptone; 0.5 % Yeast extract; 0.5 % NaCl; pH 7.0

LB-Agar

1 % Tryptone; 0.5 % Yeast extract; 0.5 % NaCl,

15 % Agar; pH 7.0

SOC-Medium

2 % Tryptone, 0.5 % Yeast extract,

0.5 % NaCl, 25 mM KCl,

100 mM MgCl

, 20 mM Glucose; pH 7.0

Ampicillin

Stock solution: 50 mg/ml in H

Working concentration: 50 µg/ml

Chloramphenicol

Stock solution: 10 mg/ml in MeOH

Working concentration: 20 µg/ml

Kanamycin

Stock solution: 50 mg/ml in H

Working concentration: 50 µg/ml

3.1.3 Buffers and solutions

Buffers and Solutions Composition

cDNA labelling

50 x aa-dNTP Mix

25 mM dATP, 25 mM dGTP, 25 mM dCTP,

15 mM dTTP, 10 mM aminoally-dUTP

1M Phosphate buffer 9.5 ml 1M K

HPO

, 0.5 ml 1M KH

; pH 8.5-8.7

Phosphate wash buffer 5 mM Phosphate buffer in 80 % Ethanol; pH 8.0

Phosphate elution buffer 4 mM Phosphate buffer; pH 8.0

Dye coupling buffer 100 mM Na

; pH 9.0 (freshly prepared)

Cy-Dyes content of one vial reconstituted in 73 µl DMSO,

(GE Healthcare) stored dessicated at -20 ° C

Stop solution

100 mM NaOAc; pH 5.2

(Dye coupling)

Microarray hybridisation

20x SSC 3 M NaCl, 0.3 M Natriumcitrate; pH 7.0

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Crosley CRSH268MW2 Bedienungsanleitung Seite 16